Patent application title: FUSION PROTEINS FOR THE TREATMENT OF CNS
Inventors:
Elliott A. Gruskin (Malvern, PA, US)
Elliott A. Gruskin (Malvern, PA, US)
Anthony O. Caggiano (Larchmont, NY, US)
Gargi Roy (Boyds, MD, US)
Jennifer Iaci (Boonton, NJ, US)
Jennifer Iaci (Boonton, NJ, US)
Michael P. Zimber (Mamroneck, NY, US)
IPC8 Class: AC12N988FI
USPC Class:
424 937
Class name: Drug, bio-affecting and body treating compositions whole live micro-organism, cell, or virus containing animal or plant cell
Publication date: 2015-10-22
Patent application number: 20150299687
Abstract:
This disclosure relates to compositions capable of use in the treatment
of spinal cord injuries and related disorders of the central nervous
system (CNS), and in particular, compositions including proteoglycan
degrading molecules and compositions capable of blocking and/or over
coming the activity of neuronal growth inhibitory molecules, as well as
fusion proteins which includes a proteoglycan degrading domain and a
domain capable of blocking and or over coming the activity of neuronal
growth inhibitory molecules.Claims:
1-45. (canceled)
46. A composition comprising: a polypeptide comprising a protein transduction domain and a proteoglycan degrading domain, wherein the proteoglycan degrading domain is selected from the group consisting of chondroitinase ABC I (SEQ ID NO: 50), chondroitinase ABC II (SEQ ID NO: 35), hyaluronidase-1 (SEQ ID NO: 38), hyaluronidase-2 (SEQ ID NO: 39), hyaluronidase-3 (SEQ ID NO: 40), hyaluronidase-4 (SEQ ID NO: 41), PH-20 (SEQ ID NO: 42), chondroitinase B (SEQ ID NO: 37), chondroitinase AC (SEQ ID NO: 36), and a combination thereof.
47. The composition of claim 46, wherein the protein transduction domain is a TAT domain.
48. The composition of claim 46, wherein the protein transduction domain and the proteoglycan degrading domain are bonded together by a linker polypeptide domain.
49. The composition of claim 46, further comprising a pharmaceutically acceptable excipient.
50. The composition of claim 46, wherein the polypeptide comprises an amino acid sequence represented by SEQ ID NO: 89.
51. The composition of claim 46, further comprising cells from the central nervous system.
52. The composition of claim 46, wherein the polypeptide further comprises a domain that promotes neural regeneration.
53. The composition of claim 52, wherein the domain that promotes neural regeneration is selected from the group consisting of L1 (SEQ ID NO: 5), a functional variant that is at least 80% L1, a functional deletion mutant of L1, GGF2 (SEQ ID NO:6), a functional variant that is at least 80% GGF2, a functional deletion mutant of GGF2, NgR27-311 (SEQ ID NO:4), a functional variant that is at least 80% NgR27-311, and a functional deletion mutant of NgR27-311.
54. The composition of claim 52, wherein the domain that promotes neural regeneration is bonded to the protein transduction domain or the proteoglycan degrading domain by a linker polypeptide domain.
55. The composition of claim 46, wherein the proteoglycan degrading domain chondroitinase ABC I (SEQ ID NO: 50) does not comprise a signal sequence.
56. A method of promoting recovery of neurological function comprising: administering to a subject in need thereof a composition comprising a polypeptide comprising a protein transduction domain and a proteoglycan degrading domain, wherein the proteoglycan degrading domain is selected from the group consisting of chondroitinase ABC I (SEQ ID NO: 50), chondroitinase ABC II (SEQ ID NO: 35), hyaluronidase-1 (SEQ ID NO: 38), hyaluronidase-2 (SEQ ID NO: 39), hyaluronidase-3 (SEQ ID NO: 40), hyaluronidase-4 (SEQ ID NO: 41), PH-20 (SEQ ID NO: 42), chondroitinase B (SEQ ID NO: 37), chondroitinase AC (SEQ ID NO: 36), and a combination thereof.
57. The method of claim 56, wherein the protein transduction domain is a TAT domain.
58. The method of claim 56, wherein the protein transduction domain and the proteoglycan degrading domain are bonded together by a linker polypeptide domain.
59. The method of claim 56, wherein the composition further comprises a pharmaceutically acceptable excipient.
60. The method of claim 56, wherein the polypeptide comprises an amino acid sequence represented by SEQ ID NO: 89.
61. The method of claim 56, wherein the composition further comprises cells from the central nervous system.
62. The method of claim 56, wherein the polypeptide further comprises a domain that promotes neural regeneration.
63. The method of claim 62, wherein the domain that promotes neural regeneration is selected from the group consisting of L1 (SEQ ID NO: 5), a functional variant that is at least 80% L1, a functional deletion mutant of L1, GGF2 (SEQ ID NO: 6), a functional variant that is at least 80% GGF2, a functional deletion mutant of GGF2, NgR27-311 (SEQ ID NO: 4), a functional variant that is at least 80% NgR27-311, and a functional deletion mutant of NgR27-311.
64. The method of claim 62, wherein the domain that promotes neural regeneration is bonded to the protein transduction domain or the proteoglycan degrading domain by a linker polypeptide domain.
65. The method of claim 56, wherein the proteoglycan degrading domain chondroitinase ABC I (SEQ ID NO: 50) does not comprise a signal sequence.
Description:
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of pending U.S. application Ser. No. 10/848,564, filed May 17, 2004, which claims the benefit and priority of U.S. Provisional Application No. 60/471,239, filed May 16, 2003, U.S. Provisional Application No. 60/471,240, filed May 16, 2003, U.S. Provisional Application No. 60/471,300, filed May 16, 2003 and U.S. Provisional Application No. 60/474,372, filed May 29, 2003. These applications are hereby incorporated by reference in their entirety.
BACKGROUND AND SUMMARY
[0002] Spinal cord injury (SCI) inflicts trauma to the cells and tissues of the central nervous system (CNS) and causes a severe and debilitating condition in the individual. Following SCI, limited regeneration of injured neurons results in permanent disability characterized by some loss of sensation, paralysis and autonomic dysfunction. One reason that neurons fail to regenerate is their inability to traverse the glial scar that develops following SCI. This glial scar contains extracellular matrix molecules including chondroitin sulfate proteoglycans (CSPGs). In vitro studies show that neurons fail to extend processes over CSPG coated surfaces, while in vivo data correlate failure of regeneration with areas of CSPG expression. Within the adult central nervous system (CNS) myelin there are also several identified axon growth inhibitor compounds like myelin associated glycoprotein (MAG), OMgp, and Reticulon 4 or Nogo that have been shown to be inhibitory for the growth of neurons.
[0003] The proteglycan degrading enzyme chondroitinase ABC type I has been used to enhance neuronal growth in a dorsal column lesion model of spinal cord injury. It has also been reported that treating a spinal cord injury with NOGO receptor antagonist promotes a certain limited degree of neuronal regeneration. It has further been reported that creating a NOGO knock out mouse resulted in certain limited and inconsistent degrees of neuronal regeneration following dorsal hemisection of the spinal cord.
[0004] Experimental treatments for injury to the CNS have utilized the application of chondroitinase to the extracellular space, however the enzyme digests CSPG in the extracellular matrix and not intracellular stores. Furthermore, diffusion of chondroitinase within the parenchyma the essential and distinctive tissue of an organ or an abnormal growth as distinguished from its supportive framework) or between anatomical compartments is limited. The limited access of drugs, imaging agents, and anesthetics, etc. to target cells and/or tissues of the Central Nervous System (CNS) may reduce the usefulness or effectiveness of any of these substances.
[0005] The delivery of therapeutic and diagnostic molecules into cells and tissues is, in part, dependent upon extracellular matrices as well as carbohydrates and proteins linked to cell membranes. The extracellular matrix is composed in part of proteoglycans, among them are the chondroitin sulfated proteoglycans (CSPGs). CSPGs are a family of proteoglycans composed of a core protein and covalently linked sulfated glycosaminoglycans. Each proteoglycan is determined by the glycosaminoglycan side chains. For CSPGs these side chains are made up of approximately 40 to 100 sulfated disaccharides composed of chondroitin 4, 6 and dermatan sulfates. The protein component of the CSPG is ribosomally synthesized and the glycosylation occurs in the endoplasmic reticulum and Golgi apparatus. The sugar chains are then sulfated at the 4 or 6 positions by several glycosaminoglycan sulfotransferases.
[0006] Transduction proteins may be used to transport polypeptides and polynucleotides cargo across anatomical barriers and into cells. For example, the TAT protein (SEQ ID NO: 2) from the human immunodeficiency virus (HIV) contains a protein transduction domain (PTD) that is involved in the transduction of HIV into cells. The PTD contains an 11 amino acid domain (TAT Peptide) (SEQ ID NO: 3) that is responsible for the activity of the PTD. The TAT Peptide (SEQ ID NO: 3) can be linked to proteins and facilitate the transduction of the proteins into cells. The mechanism of transduction is independent of the molecular weight or chemical properties of the proteins that are linked to the TAT Peptide (SEQ ID NO: 3). In vivo studies show that if a fusion protein consisting of the TAT Peptide (SEQ ID NO: 3) linked to the 120 kd enzyme, beta-galactosidase β-Gal), is injected into mice, then a robust delivery of β-Gal into a wide variety of cells is observed. When linked to the TAT transduction peptide (SEQ ID NO: 3), β-Gal activity was observed in the brain; without the TAT Peptide (SEQ ID NO: 3), β-Gal was not observed in the brain. Transport across the blood brain barrier is normally restricted to certain hydrophobic small molecules and particular low molecular weight lipophilic peptides. Transport of proteins as large as β-Gal into the brain is usually not possible without substantial disruption of the blood brain barrier, but the TAT Peptide (SEQ ID NO: 3) facilitates transport while leaving the blood brain barrier intact.
[0007] Chimeric proteins, also called fusion proteins, are hybrid proteins which combine at least parts of two or more precursor proteins or polypeptides. Chimeric proteins may be produced by recombinant technology, i.e. by fusing at least a part of the coding sequence of one gene to at least a part of the coding sequence of another gene. The fused gene may then be used to transform a suitable organism which then expresses the fusion protein.
[0008] Tat Peptide Complexes Frankel et al. (U.S. Pat. Nos. 5,804,604; 5,747,641; 5,674,980; 5,670,617; 5,652,122) discloses the use of Tat peptides to transport covalently linked biologically active cargo molecules into the cytoplasm and nuclei of cells. Frankel only discloses covalently linked cargo moieties that are (therapeutic, diagnostic or prophylactic), and does not teach or suggest the attachment of molecules that facilitate diffusion, plasticity, neurite growth, and axon regeneration. These molecules can include but are not limited to molecules that overcome neurite out growth inhibition, or promote nerve growth such as soluble NOGO antagonists like NgR27-311, neural cell adhesion molecules like L1, neurotrophic factors, growth factors, phosphodiesterase inhibitors, and inhibitors of MAG or MOG. Additionally, deletion mutants may be combined with other compounds that promote remyelination such as neuregulins (GGF2) and antibodies that promote remyelination or proteoglycan degrading molecules to Tat peptides.
[0009] Regeneration following SCI is limited because of a variety of obstacles that include the deposition of CSPG at the glial scar, demyelination of axons, lack of trophic support and lack of axonal guidance. A single therapy directed against one aspect of SCI may not be as effective as a combinatorial approach. Fusion proteins with chondroitinase will allow combinatorial therapy with a single protein. Fusion partners for chondroitinase that will be constructed in this proposal were chosen from among proteins that have evidence for efficacy in SCI.
[0010] The use of a molecule that has the ability to both degrade extracellular matrix glycoproteins and to block or overcome the inhibitory nature of myelin components may be used to improve the ability of damaged neurons to grow or regenerate compared with either treatment alone. The proteoglycan degrading molecules may also be used advantageously to provide a method of facilitating access and diffusion of substances into cells or tissues through the use of at least one enzyme capable of cleaving proteoglycans and preferably degrading chondroitin sulfate proteoglycans (CSPG).
[0011] Embodiments of the present invention include compositions that comprise polypeptides which cleave proteoglycans, polypeptides that block and/or overcome the activity of neuronal growth inhibitory molecules, or a combination of these. The compositions containing the proteoglycan degrading molecule or neuronal growth inhibitory molecules may also include molecules for transduction of the polypeptides across cell membranes and the blood brain barrier. The compositions may be used in the treatment of spinal cord injuries and related disorders of the central nervous system (CNS). The compositions can be used in the regeneration of damaged neurological tissue and facilitate the diffusion and transport of therapeutic molecules capable of blocking and/or overcoming the activity of neuronal growth inhibitory molecules into damaged or diseased tissue.
[0012] Embodiments of the present invention include compositions and methods for their use to facilitate delivery and diffusion of therapeutics or diagnostic agents, and preferably agents that promote regeneration of nerves and axons, into cells or tissues. Preferably the composition includes the use of an enzyme capable of cleaving chondroitin sulfate proteoglycans (CSPG) to increase the diffusion of these agents into cells or tissues of the central nervous system.
[0013] Compositions of the present invention may include chimeric or fusion proteins capable of systemic use in the treatment of spinal cord injuries and related disorders of the central nervous system (CNS), and in particular, fusion proteins capable of crossing the blood brain barrier. The fusion protein may include a polypeptide transduction domain, a polypeptide domain capable of degrading a proteoglycan, preferably a domain cleaving chondroitin sulfate proteoglycan (CSPG), a polypeptide domain that blocks and or over comes the activity of neuronal growth inhibitory molecules, or any combination of these polypeptide domains that may be used in the treatment of spinal cord injuries and related disorders of the central nervous system (CNS). The various polypeptide domains may be linked or chemically bonded together by polypeptide linkers.
[0014] Compositions of the present invention include polynucleotides which encode for the chimeric or fusion proteins capable of systemic use in the treatment of spinal cord injuries and related disorders of the central nervous system (CNS), and in particular, they encode for fusion proteins capable of crossing the blood brain barrier. The polynucleotides which encode for these chimeric or fusion proteins may include a polynucleotide domain encoding for a polypeptide transduction domain, a polynucleotide domain encoding for a polypeptide domain capable of degrading a proteoglycan, preferably cleaving chondroitin sulfate proteoglycan (CSPG), a polynucleotide domain encoding for a polypeptide domain that blocks and or over comes the activity of neuronal growth inhibitory molecules, or any combination of these domains that may be used in the treatment of spinal cord injuries and related disorders of the central nervous system (CNS). The polynucleotide also includes one or more polynucleotide domains that encode for polypeptides that link the domains of the polypeptide together to form the fusion protein.
[0015] One embodiment of the present invention is a composition and a method for its use that facilitates the access and distribution of therapeutic and diagnostic agent in the composition into cells, through membranes or into tissues by the use of composition that includes at least one enzyme capable of cleaving proteoglycans, preferably the composition includes a fusion protein having an enzyme capable of cleaving CSPGs. The molecules or agents in the composition may include one or more of Growth factors including, Brain Derived Neurotrophic Factor, Insulin-like Growth Factor, Fibroblast Growth Factor, Ciliary Neurotrophic Factor, Glial Derived Neurotrophic Factor, Transforming Growth Factor, Glial Growth Factor 2, L1, GM1, Vascular Endothelial Growth Factor, Nerve Growth Factor, Immunophilins. Molecules in the composition can include fluorescent or contrast agents for imaging. The agents may include cells for transplant--stem cells, neurons, others, cells as delivery agents, chemotherapeutic agents, antibiotics, antibody therapies, Nogo receptor antagonists, other chondroitinase enzymes. The composition may include a transduction domain, an enzyme capable of cleaving proteoglycans, or both. Preferably the composition includes a fusion protein having a transduction domain, an enzyme domain capable of cleaving proteoglycans, or both. The fusion protein can facilitate the transport or modifies transport of such agents into cells, tissues, and/or otherwise inaccessible locations; and/or to enhance penetration rates, distance of penetration; or provide more even concentration distribution. Preferably the modified transport occurs through the use of at least one enzyme capable of cleaving CSPGs. The compositions can be used for treating a CNS injury, preferably the composition is used in the treatment of neuronal damage from a contusion injury.
[0016] Embodiments of the present invention include chimeric proteins of a proteoglycan degrading domain linked to a polypeptide that blocks the action of neuronal growth inhibitors such as but not limited to a Nogo-receptor antagonist (NgR27-311) domain or variant linked to a chondroitinase like chondroitinase ABC I or a variant of chondroitinase having one or more N terminal amino acids deleted. The compound may include chimeric proteins of a proteoglycan degrading domain linked to a polypeptide that is a neural cell adhesion promoter such as an L1 neural cell adhesion promoter domain or variant linked to chondroitinase ABC I or a variant of chondroitinase having one or more N terminal amino acids deleted. The chimeric proteins may include chimeric proteins of a proteoglycan degrading domain linked to a polypeptide that is a glial cell stimulator, such as but not limited to a GGF2 glial cell stimulator or variant linked to chondroitinase ABCI or a variant of chondroitinase having one or more N terminal amino acids deleted.
[0017] An E. Coli recombinant expression system and purification process can be used to produce essentially pure and catalytically active chondroitinase ABCI. These methods may be modified for producing chimeras of proteoglycan degrading molecules and other agents.
[0018] The chimera may be assayed for chondroitinase enzymatic activity and the specific biological activity of each fusion partner. Methods to measure the activities of the chimera may be modified for those used to measure chondroitinase activity including a spectrophotometric assay, zymography, an HPLC assay to detect CSPG disaccharide digestion products and an in vito neurite outgrowth assay. A neuron growth cone collapse assay can be used to evaluate NOGO receptor antagonists and a neurite outgrowth assay can be used measure L1 activity. GGF2 activity may be measured using a Schwann cell proliferation assay.
[0019] The compositions and method of the present invention can be used for the treatment of spinal cord injuries and in the promotion of regeneration of axons. The compositions of the present invention can also be used to promote plasticity, regrowth, repair, and/or regeneration of dysfunctional neurons in the CNS that have been damaged as a result of disease, such as degenerative diseases including Alzheimer's and Parkinson's disease. Advantageously, the use of proteoglycan degrading polypeptides or membrane transducing polypeptides in the compositions of the present invention also promote diffusion and access of damage or diseased tissue to other therapeutic agents promoting the regeneration of neurons.
DESCRIPTION OF THE DRAWINGS
[0020] The file of this patent contains at least one drawing/photograph executed in color. Copies of this patent with color drawing(s)/photograph(s) will be provided by the Office upon request and payment of the necessary fee.
[0021] In part, other aspects, features, benefits and advantages of the embodiments of the present invention will be apparent with regard to the following description, appended claims and accompanying drawings where:
[0022] FIG. 1 is an illustrative example of a TAT-chondroitinase ABCI construct; the DNA sequence for the entire gene fragment fused with the Tat sequence followed by the 5-glycine linker (SEQ ID NO: 45); the sense and antisense oligonucleotides having the sequences as 5'-tatgtatggtc gtaaaaagcgtc gtcaacgtcgtcgtgg tggtggtggtca-3' (SEQ ID NO: 92) and 5'-tatgaccaccaccaccaccacgac gacgttgacgac gcttttt acgaccataca-3' (SEQ ID NO: 93) were annealed and ligated at the NdeI site which is at the 5' end [f]of ABCI gene cloned in pET15b (Novagen) at the NdeI and BamHI sites.
[0023] FIG. 2 are Brain section images; (I) illustrating lobes from adult rat which were incubated in beta-galactodidase alone (B&D), or with the addition of Choindroitinase ABCI (A, 0.5 U/ml or C, 0.005 U/ml); (II) Eosin Y penetration through the cortex of a Choindroitinase treated brain hemisphere and control showing approximately the same penetration, Eosin Y is a is a zwitterionic, having an overall negative charge at the low pH it was used at, and it is 692 kDa; (III)A saturate solution of Congo Red demonstrates greater penetration through the cortex of a Chondroitinase treated brain hemisphere as compared to untreated brain, Congo red is a negatively charged dye of 697 kDa;
[0024] FIG. 3 (A) is a diagram representing the full length GGF protein (GGF2-M1-E422) and the three GGF2 fragments containing the Ig and EGF domains GGF2-L250-C402, Ig and EGF domains GGF2-L250-E422 to C-terminal; and EGF domains GGF2-T350-C402; (B) a schematic presentation of chimeric proteins of a proteoglycan degrading molecule like chondroitinase ABCI and a neuregulin 1 gene isoform GGF2 such as N-terminal fusion chimera between chondroitinase ABCI-NΔ60-CΔ80 and GGF2-FL, between chondroitinase ABCI-NΔ60-CΔ80 and GGF2-L250-C402, between chondroitinase ABCI-NΔ60-CΔ80 and GGF2-L250-E422, and between chondroitinase ABCI-NΔ60-CΔ80 and GGF2-T350-C402, and C-terminal fusion chimera (lower) such as between chondroitinase ABCI-FL and GGF2-FL, between chondroitinase ABCI-FL and GGF2-L250-C402, between chondroitinase ABCI-FL and GGF2-L250-E422, and between chondroitinase ABCI-FL and GGF2-T350-C402.
[0025] FIG. 4 illustrate the structure of (A) an NgR27-311--N terminal chondroitinase ABCI chimeric protein without, and with, a peptide spacer or linking group; (B) an NgR27-311--C terminal chondroitinase ABCI chimeric protein without, and with, a peptide spacer or linking group; (C) an extracellular domain L1 N-terminal chondroitinase ABCI chimeric protein without, and with, a spacer or linking peptide; (D) an extracellular domain L1 C-terminal chondroitinase ABCI chimeric protein without, and with, a spacer or linking peptide.
[0026] FIG. 5 (A)BBB scores from spinal cord injured animals treated with chondroitinase (Chondroitinase ABC I), penicillinase or artificial cerebrospinal fluid (aCSF); (B) Parasagittal sections of spinal cords from injured animals treated with chondroitinase (left column) or penicillinase (right column). Sections were cut at 30 microns. Each set of images contains four parasagittal sections realigned with the rostral cord to the left and the caudal cord to the right. The most central section of each set has been removed and replaced below to aid visualization. Pairs of images are Weil stained, anti-GFAP and an amino cupric stain of neuronal degeneration (top to bottom). (C) and (D) Distribution of individual BBB scores according to treatment group. Scores are BBB scores at ten weeks post surgery. Group averages are shown below the data points.
DETAILED DESCRIPTION
[0027] Before the present compositions and methods are described, it is to be understood that this invention is not limited to the particular molecules, compositions, methodologies or protocols described, as these may vary. It is also to be understood that the terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.
[0028] It must also be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to a "cell" is a reference to one or more cells and equivalents thereof known to those skilled in the art, and so forth. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, the preferred methods, devices, and materials are now described. All publications mentioned herein are incorporated by reference. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.
[0029] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event occurs and instances where it does not.
[0030] Suitable enzymes that cleave CSPGs include chondroitinase ABC type I, chondroitinase ABC Type II, chondroitinase AC, and chondroitinase B, or mammalian enzymes with chondroitinase-like activity such as hyaluronidase-1, hyaluronidase 2, hyaluronidase 3, hyaluronidase 4, cathepsins, ADAMTs and PH-20, or mixtures thereof.
[0031] CSPGs are a family of proteoglycans composed of a core protein and covalently linked sulfated glycosaminoglycans. The polysaccharide chains are cleaved by several enzymes, including a family of chondroitinases. To date, this enzyme family contains at least four members, Chondroitinase ABC I, ABC II, AC and B. Chondroitinase ABC I is an exo-lyase which cleaves both chondroitin and dermatan sulfates. It has been used extensively in the study of in vitro neuronal regeneration over CSPG-laden substrates and more recently in in vivo studies following CNS injury.
[0032] Proteins and polypeptide that may be used in the compositions and fusion proteins of the present invention are those which promote plasticity as well as the regeneration of injured or diseased neurons and axons. These regenerating proteins and polypeptides may include cell adhesion proteins, those which stimulate glial cells, and polypeptides which block the inhibitory effect of proteins that act as axon growth inhibitors.
[0033] Plasticity of the nervous system refers to any type of functional reorganization. This reorganization occurs with development, learning and memory and brain repair. The structural changes that occur with plasticity may include synapse formation, synapse removal, neurite sprouting and may even include strengthening or weakening existing synapses. Regeneration is generally differentiated from plasticity by the long range growth of axons in disrupted tracts that is characteristic of regeneration.
[0034] Proteins and polypeptides are capable of blocking the activity of neuronal growth inhibitory molecules may include peptides and polypeptides that block the inhibitory properties of peptide such as but not limited Nogo, MAG, and OMgp. Suitable compositions that overcome the activity of neuronal growth inhibitory molecules include but are not limited to Protein Kinase C family inhibitors, Rho Kinase family inhibitors and agents, such as phosphodiesterase inhibitors, that increase intracellular cyclic AMP, and L1. The (NgR27-311) peptide has been shown to inhibit binding of Nogo66, OMgp, MAG, and MOG to membrane-bound NogoR and overcome the inhibitory effects of Nogo on the regeneration of nerve processes.
[0035] Proteins and polypeptides that affect cell adhesion or stimulate cells may include but are not limited to poly peptides such as L1 and GGF2. L1 is a neural cell adhesion protein that is a potent stimulator of neurite outgrowth in vitro for which has been found that treatment of acute SCI in rodents using a soluble form of L1 leads to an increase in the recovery of neurological function. GGF2 stimulates glial cells and has been shown to improve clinical outcome measures in a murine model of experimental allergic encephalomyelitis (EAE) likely as a result of the stimulation of oligodendrocytes to promote remyelination.
[0036] Cell membrane-permeant peptide sequences useful in practicing the present invention include, but are not limited to, RQARRNRRRRWRERQR-51 (HIV-1 Rev protein basic motif; (SEQ ID NO:43)); MPKTRRRPRRSQRKRPPTP-119 (HTLV-1 Rex protein basic motif; (SEQ ID NO:44)) (Kubota et al. 1989); the third helix of the homeodomain of Antennapedia (Derossi, et al., J. Biol. Chem. 271:18188-93, 1996) (43-RQIKIWFQNRRMKWKK-58 (SEQ ID NO:45)); a peptide derivable from the heavy chain variable region of an anti-DNA monoclonal antibody (Avrameas, et al., Proc. Natl. Acad. Sci. 95:5601-06, 1998) (VAYISRGGVSTYYSDTVKGRFTRQKYNKRA (SEQ ID NO:46)); and the Herpes simplex virus VP22 protein (Elliot and O'Hare, Cell, 88:223-33, 1997) (1-MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQR GEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPP AGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAP ASRAPTVQLWQMSRPRTDEDLNELLGITHRVTVCEGKNLLQRANELVNPDV VQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE-246 (SEQ ID NO:47)). In a preferred embodiment, the basic peptide is derivable from the human immunodeficiency virus type 1 (HIV-1) Tat protein (Fawell et al., Proc. Natl. Acad. Sci., 91:664-68, 1994). In particular, the Tat peptide can comprise any sequential residues of the Tat protein basic peptide motif 37-72 (Vives et al. 1997) (37-CFITKALGISYGRKKRRQRRRPPQGSQTHQVSLSKQ-72 (SEQ ID NO:48)). The minimum number of amino acid residues can be in the range of from about three to about six, preferably from about three to about five, and most preferably about four, i.e., the minimal requirement for one alpha helical turn. A preferred embodiment comprises Tat protein residues 48-57 (GRKKRRQRRR) (SEQ ID NO:49).
[0037] Suitable PTD domains include those derived from the TAT protein (SEQ ID NO: 2). Tat proteins and peptides: Tat (SEQ ID NO: 2) is an 86-amino acid protein involved in the replication of human immunodeficiency virus type 1 (HIV-1). The HIV-1 Tat transactivation protein (SEQ ID NO: 2) is efficiently taken up by cells (Mann and Frankel 1991; Vives et al. 1994), and low concentrations (nM) are sufficient to transactivate a reporter gene expressed from the HIV-1 promoter (Mann and Frankel 1991). Exogenous Tat protein (SEQ ID NO: 2) is able to translocate through the plasma membrane and reach the nucleus to transactivate the viral genome.
[0038] A region of the Tat protein (SEQ ID NO: 2) centered on a cluster of basic amino acids is believed to be responsible for this translocation activity (Vives et al. 1997). Tat peptide-mediated cellular uptake and nuclear translocation have been demonstrated in several systems (Vives, et al., J Biol Chem 272:16010-16017, 1997; Jones, Genes Dev 11:2593-2599, 1997). Chemically coupling a Tat-derived peptide (residues 37-72) (SEQ ID NO:48) to several proteins results in their internalization in several cell lines or tissues (Fawell, et al., Proc Natl Acad Sci USA 91:664-668, 1994; Anderson, et al., Biochem Biophys Res Commun 194:876-8884, 1993; Fahraeus, et al., Curr Biol 6:84-91, 1996; Nagahara, et al., Nat Med 4:1449-1452, 1998). A synthetic peptide consisting of the Tat basic amino acids 48-60 with a cysteine residue at the C-terminus coupled to fluorescein maleimide translocates to the cell nucleus as determined by fluorescence microscopy (Vives et al. 1997). In addition, a fusion protein (Tat-NLS-β-Gal) consisting of Tat amino acids 48-59 fused by their amino-terminus to β-galactosidase amino acids 9-1023 translocates to the cell nucleus in an ATP-dependent, cytosolic factor-independent manner (Efthymiadis et al. 1998).
[0039] Chimeric proteins, also referred to in the art as fusion proteins, are hybrid proteins which combine at least parts of two or more precursor proteins or peptides. Chimeric proteins may be produced by recombinant technology, i.e. by fusing at least a part of the coding sequence of one gene to at least a part of the coding sequence of another gene. Where desirable, one or more genes for linker peptides may be fused to the coding sequence of genes for the other polypeptide domains in the fusion protein. The fused gene may then be used to transform a suitable organism such as but not limited to E. coli or CHO cells which then expresses the fusion protein.
[0040] Genes encoding either N or C terminal deletion mutants of polypeptide domains of the fusion proteins can be used in constructs for expression of the fusion proteins. Preferably the generated deletion mutants maintain their catalytic proteoglycan degrading activity, blocking activity, growth activity, or transduction activity. Generated deletion mutants of the proteoglycan degrading molecules like chondroitinase ABCI enzyme where the mutant is missing a certain number of amino acids from the N and or C-terminal are those that retain some proteoglycan degrading activity. N-terminal deletions of chondriotinase like chondroitinase ABC I maintain a histidine-tag that is attached to the N-terminus. It is expected that a TAT-deletion mutant chondroitinase ABC I fusion DNA construct can be expressed without removal of the TAT polypeptide during expression. For example a TAT peptide can be fused at the N-terminus of either ABCI-NΔ20 or ABCI-NΔ60 deletion mutant. Fragments of polypeptides, such as those shown in FIG. 3 for GGF2, may be used in the construction of chimeric fusion proteins.
[0041] Catalytically active deletion mutants of chondroitinase ABCI can be prepared for example but not limited to deleting 20, 40 and 60 amino acids respectively from the N-terminus of the mature ABCI protein. Deletion of 60 amino acids from the N terminal and 80 amino acids from the C-terminal end may also be used to make a deletion mutants of a proteoglycan degrading chondroitinase ABCI. These deletion mutants and those of other proteoglycan degrading molecules may be used for construction of N-terminal fusion chimeric protein. Detailed comparative biochemical studies can be done to determine the efficacy of mature chondroitinase ABCI versus various deletion mutant in compositions and fusion proteins with respect to the substrate specificity, substrate binding and tissue penetration.
[0042] A mutant of chondroitinase ABCI that has native protein structure, but lacks catalytic activity may be prepared as a null or a negative control for bioassays and SCI studies. Based on the crystal structure of chondroitinase ABCI a site-specific mutant designated H501a and Y508a to knock out catalytic activity in the putative active site can be prepared. Such mutants can be tested for inactivation of catalytic activity and SEC to compare to the wild-type enzyme. If the null activity mutant is successfully created it will provide a negative control for the various fusion proteins for use in bioassays and ultimately in SCI animal studies.
[0043] An E. coli expression system may be used to make chondroitinase using PET expression vectors (Novagen). The GGF2-chondroitinase ABCI fusion protein may be expressed in E. coli. Constructs for Tat-chondroitinase deletion mutant fusion proteins, Tat-GGF2 fusion proteins, or Tat-chondroitinase-GGF2 fusion proteins may be expressed from E. coli. Other fusion proteins can be expressed in CHO cell lines.
[0044] Table 1 illustrates various non-limiting components which may be used in compositions of the present invention as a mixture, and preferably as a fusion or chimeric molecule. The composition or chimeric molecule described may include one or more of the molecules in Table 1. In the case of chimeric molecules one or more linker segments, preferably polypeptides are used.
TABLE-US-00001 TABLE 1 Components of Compositions Proteoglycan degrading Therapeutic, diagnostic, Transduction molecules molecules receptor antagonist molecules Tat protein residues 48-57 Any agent that degrades Any peptide that blocks the (SEQ ID NO: 49) extracellular matrix inhibitory properties of Nogo, HIV-1 Rev protein basic glycoproteins including: MAG, OMgp including: motif (SEQ ID NO: 43) Chrondroitinase ABC I, Nogo peptide 1-40 HIV-1 Rev protein basic Chondroitinase ABC II, Any component of Nogo motif(SEQ ID NO: 43) Chondroitinase AC, peptide 1-40 that maintains the Herpes simplex virus Chondroitinase B, ability to block the inhibitory VP22 protein Hyaluronidase 1, properties of Nogo Hyaluronidase 2, Other blocking peptides of Hyaluronidase 3, Nogo Hyaluronidase 4, Antibodies that recognize Nogo PH20 Blocking peptides of MAG deletion and or substitution Antibodies that recognize MAG mutants of the above listed Blocking peptides of OMgp molecules that maintains Antibodies that recognize enzymatic activity. OMgp Antibodies that recognize the Nogo-66 receptor Blocking peptides of the p75 receptor Antibodies that recognize the p75 neurotrophin receptor Peptides or antibodies that block other receptors of Nogo, MAG and/or OMgp Peptide that overcomes the inhibitory properties of Myelin, Nogo, MAG, OMgp including: Protein Kinase C family inhibitors* Rho Kinase family inhibitors Agents that increase intracellular cAMP concentration L1
[0045] A schematic illustration of non-limiting versions of chimeric fusion proteins of the present invention are illustrated in FIG. 4A and FIG. 4B.
[0046] A peptide component from any column in Table 1 could be linked by an oligopeptide linker that is well known in the art. A glycine rich peptide, for example Gly-Gly-Gly-Gly-Gly (SEQ ID NO: 56), or linkers prepared including any of the naturally occurring amino acids as well as substituted or beta or gamma amino acids like 4-aminobutyric acid or 6-aminocaproic acid can be used. Other linkers including but not limited to alkyl diamines, amino, or alkyl diols may also be used. Preferably the transduction component of the fusion protein is in a terminal position in the polypeptide. Other examples of common linkers may include but would not be limited to Gly-Gly-Ala-Gly-Gly (SEQ ID NO: 57), Gly/Ser rich linkers (for example Gly4Ser3 (SEQ ID NO: 94)), or Gly/Ala rich linkers. Additionally, linkers may be of any length and design to promote or restrict the mobility of components in the fusion protein.
[0047] The incorporation of non-natural amino acids, including synthetic non-native amino acids, substituted amino acids, or one or more D-amino acids into the peptides (or other components of the complexes) of the present invention (subsequently referred to herein as "D-peptides") is advantageous in a number of different ways. D-amino acid-containing peptides exhibit increased stability in vitro or in vivo compared to L-amino acid-containing counterparts. Thus, the construction of peptides incorporating D-amino acids can be particularly useful when greater intracellular stability is desired or required. More specifically, D-peptides are resistant to endogenous peptidases and proteases, thereby providing better oral transepithelial and transdermal delivery of linked drugs and conjugates, improved bioavailability of membrane-permeant complexes, and prolonged intravascular and interstitial lifetimes when such properties are desirable. The use of D-peptides can also enhance transdermal and oral transepithelial delivery of linked drugs and other cargo molecules.
[0048] In a spinal cord injury, the axons of ascending sensory and descending motor neurons are disrupted, that can result in the loss of sensation and paralysis. These axons fail to regenerate successfully leading to permanent disability. A scar envelopes the site of the injury which is believed to wall off the area of fragile tissue, stabilize the blood brain barrier, and prevent an overwhelming cascade of uncontrolled tissue damage. This scar is composed of hypertrophic glial cells and an extracellular matrix (ECM). Chondroitin sulfate proteoglycans (CSPGs) are one important component of the scar. They are expressed by glial cells and deposited in the ECM in regions of blood brain barrier breakdown. In vitro evidence demonstrates that these CSPGs are potently inhibitory for the growth of axons and without wishing to be bound by theory, are believed to contribute to the failure of the spinal cord axons to regenerate and reform functional synapses. In vivo studies have demonstrated that regenerating axons are able to grow into and even beyond the scar.
[0049] CSPGs and white matter components are generally accepted as molecular barriers that neurons must overcome in order to regenerate and reestablish functional connections after injury. Transplanted adult sensory neurons placed distal to a forming scar can regenerate robustly even along degenerating white matter pathways, however, regeneration ceases abruptly as axons enter the proteoglycan containing glial scar. Treatment of CNS white matter pathways with chondroitinase enhances the ability of neurons to grow in these substrates.
[0050] Central nervous system tissues are tightly compacted with cells and have limited extracelluar space. The proteins and carbohydrates of the extracellular matrix provide charge and osmotic forces as well as specific and non-specific binding sites which may prevent the penetration of therapeutic agents. The enzymatic cleavage of these matrix and cellular components may later or facilitate the access of compounds or cells through tissues. A proteoglycan degrading molecule like Chondroitinase ABC I that is an enzyme that digests chondroitin sulfate proteoglycans can be used to promote diffusion of therapeutic molecules into the CNS. Tat peptides transport covalently linked biologically active cargo molecules into the cytoplasm and nuclei of cells. In the case of a fusion protein having a protein transduction polypeptide domain like the HIV tat protein (SEQ ID NO: 3), therapeutic molecules for axon regeneration may be delivered across the blood brain barrier.
[0051] Treatment of SCI model injuries, preferably contusion model injury, can be used to determine the degree of regeneration and functional recovery achieved by compositions and method of the present invention. The degree of functional recovery can be demonstrated by improved corticospinal tract conduction, improved tape removal, beam walking, grid walking and paw placement following chondroitinase treatment of a dorsal column lesion. Motor skill improvement as well as autonomic function: bowel, bladder, sensor and sexual function may also be used as measures of function improvement and related to molecular structure and components in the compositions of the present invention.
[0052] In addition to the ability of chondroitinase to enhance regeneration, chondroitinase digests components of the perineuronal network (PNN). The density of the PNN is variable within the CNS and is particularly dense in the somatosensory, auditory, visual cortices and the hippocampus. PNN has also been demonstrated to be dense around spinal motor neurons. The inventors have discovered the dense PNN within the dorsal horn of the cord. Digestion of the PNN can enhance plasticity within the hippocampus and visual cortex. Plasticity within intact systems in incomplete SCI, especially in the region of the central pattern generators or in the reticular core, may support the function of damaged or destroyed systems. Promotion of plasticity in these systems may be one mechanism other than or in addition to regeneration by which chondroitinase can improve function following CNS injury. Furthermore, regeneration and plasticity may work in concert to affect recovery following injury; indeed, the corticospinal tract has been shown to be critical for modulation of spinal cord plasticity.
[0053] Recovery of neurological function following contusion injury in the CNS or a disease state may be promoted by administering the fusion proteins or mixtures including one or more of the components in Table 1, to cells, a tissue, or a subject having damaged or diseased neurons whether the injury or disease is immediate or long-standing.
[0054] The fusion proteins herein are administered in an amount effective to degrade CSPGs and thereby promote the recovery of neurological function. Once the proteins or polypeptides in the compositions have been purified to the extent desired, they may be suspended or diluted in an appropriate physiological carrier or excipient for SCI treatment. In models of SCI, effective intrathecal doses in rats have been about 0.06 units on alternate days for 14 days. A dose for a 70 kilogram human may be about 17 Units. At about 100 Units/milligram, this would equal about 170 micrograms. Doses of up to 20 Units appear safe in the rat. Compositions including a proteoglycan degrading molecule in a mixture or as part of a fusion protein diluted in a carrier or pharmaceutically acceptable excipient can be injected, generally at concentrations in the range of 1 μg to 500 mg/kg of host. Administering the agent can be by bolus injection, intravenous delivery, continuous infusion, sustained release from implants, or sustained release pharmaceuticals. Administration may be by injection, such as intramuscularly, peritoneally, subcutaneously, intravenously. Oral administration may include tablets or capsules, preferably the oral dosage is a sustained release formulation for once or twice daily administration. Percutneous administration can be once per day, and is preferably less than once per day administration. Administration to the human patient or other mammalian subject may be continued until a measurable improvement in autonomic or motor function in the patient is achieved.
[0055] The chondroitinase PTD fusion proteins can be administered with a suitable pharmaceutical carrier. The administration of the compositions of the present invention as mixtures or chimeric proteins can be topical, local or systemic. The chimeric fusion proteins may also be secreted by genetically cells, preferably a chimeric fusion protein having a proteoglycan degrading portion like chondroitinase and a transduction polypeptide portion like TAT can be secreted by genetically modified cells that are implanted, either free or in a capsule, at or near the site of CNS injury.
[0056] Once the compositions either as mixtures or fusion proteins are administered, degradation of CSPGs removes the inhibitory molecules that block neurite outgrowth, and allow the regeneration of neurites into the affected area. For example, the chondroitinase AC and chondroitinase B degrade CS and DS, respectively, resulting in unsaturated sulfated disaccharides. Chondroitinase AC cleaves CS at 1, 4 glycosidic linkages between N-acetylgalactosamine and glucuronic acid in the polysaccharide backbone of CS. Cleavage occurs through beta-elimination in a random endolytic action pattern. Chondroitinase B cleaves the 1, 4 galactosamine iduronic acid linkage in the polysaccharide backbone of DS. The cleavage of both CS and DS occurs through a beta-elimination process which differentiates these enzymatic mechanisms from mammalian GAG degrading enzymes. Chondroitinase ABCI, and chondroitinase ABCII, are exo and endo lyases that cleave both CS and DS. The removal of CS and DS from the glial scar permits the regeneration of neurite outgrowths into the injured area.
[0057] Mixtures of any of these fusion polypeptides may be used to provide a therapeutic treatment for CNS injuries and disorders which may include but are not limited to contusion injury, traumatic brain injury, stroke, multiple sclerosis, brachial plexus injury, amblioplia, spinal cord injuries. Spinal cord injuries includes disease and traumatic injuries, such as the crushing of neurons brought about by an auto accident, fall, contusion, or bullet wound, as well as other injuries. Practice of the present methods can confer clinical benefits to the treated mammal, providing clinically relevant improvements in at least one of the subject's motor coordination functions and sensory perception. Clinically relevant improvements can range from a detectable improvement to a complete restoration of an impaired or lost function of the CNS.
[0058] The regeneration of the nerve cells into the affected CNS area allows the return of motor and sensory function. Clinically relevant improvement will range from a detectable improvement to a complete restoration of an impaired or lost nervous function, varying with the individual patients and injuries.
[0059] A variant of a protein or fragments thereof refer to a molecule substantially similar to either the entire protein or a fragment, which possesses biological activity that is substantially similar to a biological activity of the complement protein or fragments. A molecule is substantially similar to another molecule if both molecules have substantially similar structures or if both molecules possess a similar biological activity.
[0060] Variants of complement proteins or fragments thereof are produced by chemical or recombinant means. Variants of the polynucleotides constructed to express fusion proteins may also be made. The variants may include, for example, deletions from, or insertions or substitutions of, amino acid residues within the amino acid sequence, or deletion, substitution, or insertion of nucleic acids from a sequence encoding for a particular fusion protein or polypeptide domain in the fusion protein. For example, in some cases the removal of one or more amino acid residues from a chondroitinase polypeptide can be made without significant change in its CSPG degradation activity. Substantial changes in functional properties like proteoglycan degradation or blocking activity against axon growth inhibitors are made by selecting substitutions that are less conservative, ie. that differ more significantly in their effect on maintaining the structure, charge or hydrophobicity of the peptide backbone in the area of the substitution.
[0061] Most deletions, insertions, and substitutions are not expected to produce radical changes in the characteristics of the protein molecule; however, when it is difficult to predict the exact effect of the substitution, deletion, or insertion in advance of doing so, one skilled in the art will appreciate that the effect will be evaluated by routine screening assays. For example, a change in the axon regeneration character of the polypeptide molecule, through more or less proteoglycan degradation can be measured with functional recovery tests as well as HPLC assays to detect CSPG disaccharide digestion products.
[0062] The present invention relates to the use of a proteoglycan degrading molecule, an HIV tat protein, or a tat-derived polypeptide, or a combination of these as a mixture or fusion protein to deliver a molecule of interest into the CNS or a site in the CNS where neuronal tissue damage has occurred by disease or trauma. In particular the damage site in the CNS is where scaring has occurred as a result of a contusion injury. The molecule of interest, optionally referred to as an agent or cargo molecule can be a therapeutic molecule which promotes plasticity, axon growth, a diagnostic molecule, or a proteoglycan degrading molecule. In the case of a fusion protein having a protein transduction polypeptide domain like the HIV tat protein (SEQ ID NO: 3), therapeutic molecules for axon regeneration may be delivered across the blood brain barrier or proteoglycan degrading molecule can be delivered into cells to degrade cellular stores of proteoclycans and promote axon regeneration.
[0063] Transport polypeptides of this invention may be advantageously attached to cargo molecules by chemical cross-linking or by genetic fusion. A unique terminal cysteine residue is a preferred means of chemical cross-linking According to some preferred embodiments of this invention, the carboxy terminus of the transport moiety is genetically fused to the amino terminus of the cargo moiety. Embodiment of the present invention consists of an amino-terminal methionine followed by tat residues 47-58 (SEQ ID NO: 49), followed by a chondroitinase polypeptide.
[0064] It will be appreciated that the entire 86 amino acids which make up the tat protein may not be required for the uptake activity of tat. For example, a protein fragment or a peptide which has fewer than the 86 amino acids, but which exhibits uptake into cells and or can cross the blood brain barrier, can be used (a functionally effective fragment or portion of tat). For example, tat protein containing residues 1-72 can be sufficient for uptake activity and tat residues 1-67 can mediate the entry of a heterologous protein into cells. Synthetic peptide containing tat residues 1-58 can have uptake activity.
[0065] The tat peptide can be a single (i.e., continuous) amino acid sequence present in tat protein or it can be two or more amino acid sequences which are present in tat protein, but in the naturally-occurring protein are separated by other amino acid sequences. As used herein, tat protein includes a naturally-occurring amino acid sequence which is the same as that of naturally-occurring tat protein, its functional equivalent or functionally equivalent fragments thereof (peptides). Such functional equivalents or functionally equivalent fragments can possess uptake activity into the cell or across the blood brain barrier that is substantially similar to that of naturally-occurring tat protein. Tat protein can be obtained from naturally-occurring sources or can be produced using genetic engineering techniques or chemical synthesis.
[0066] The amino acid sequence of naturally-occurring HIV tat protein (SEQ ID NO: 2) can be modified, by addition, deletion and/or substitution of at least one amino acid present in the naturally-occurring tat protein, to produce modified tat protein (also referred to herein as tat protein or polypeptide). Modified tat protein or tat peptide analogs with increased stability can thus be produced using known techniques. Therefore, tat proteins or peptides may have amino acid sequences which are substantially similar, although not identical, to that of naturally-occurring tat protein or portions thereof. In addition, cholesterol or other lipid derivatives can be added to tat protein to produce a modified tat having increased membrane solubility.
[0067] Naturally-occurring HIV-1 tat protein (SEQ ID NO: 2) has a region (amino acids 22-37) wherein 7 out of 16 amino acids are cysteine. Those cysteine residues are capable of forming disulfide bonds with each other, with cysteine residues in the cysteine-rich region of other tat protein molecules and with cysteine residues in a cargo protein or the cargo moiety of a conjugate. Such disulfide bond formation can cause loss of the cargo's biological activity. Furthermore, even if there is no potential for disulfide bonding to the cargo moiety (for example, when the cargo protein has no cysteine residues), disulfide bond formation between transport polypeptides can lead to aggregation and insolubility of the transport polypeptide, the transport polypeptide-cargo conjugate, or both. The tat cysteine-rich region may be deleted to avoid disulfide bond formation and prevent aggregation and insolubility of the transport polypeptide, the transport polypeptide-cargo conjugate, or both.
[0068] Chondroitinase is also able to promote plasticity in regions of the CNS with significantly dense PNN, including cortex, tectum, hippocampus and spinal cord. It is reasonable that some combination of effects, including regeneration, sprouting and plasticity are responsible for the improvement in function following SCI with chondroitinase treatment or treatment with fusion molecules including chondroitinase or other proteoglycan degrading molecule.
[0069] NbR27-311: Nogo is a high molecular weight myelin component that inhibits neurite outgrowth. The amino terminal region (Nogo66) is the part of the molecule that is specifically associated with the inhibition of neurite outgrowth. Expression cloning methods revealed the receptor for Nogo66 (NgR) is a GPI anchored glycoprotein expressed mainly on neurons. NgR interacts not only with Nogo, but also with other myelin associated inhibitors such as MAG and MOG. Due to its central role in the inhibitory properties of myelin on neurons, NgR has been the target for approaches to antagonize its interactions with its ligands. The soluble segment of NgR interacts with Nogo66, MAG and MOG. The region spans residues 27-311 and this NgR fragment can therefore act as a decoy receptor that will interfere with myelin associated inhibition of neurons. If NgR27-311 is linked to proteoglycan degrading molecule like chondroitinase ABCI to form a chimeric fusion protein, the NgR27-311 polypeptide domain of the fusion protein would be expect to limit the myelin-associated inhibition of neurite outgrowth and promote axonal regeneration in chondroitinase digested regions of a spinal cord injury.
[0070] For cloned (NgR27-311), the precise region of interest, or fragments can be derived from the initial clone. The biological activity of NgR27-311, its fragments, and fusion polypeptides including NgR27-311 may be confirmed using an in vitro assay for the collapse of growth cones on neurons. The growth cone collapse assay has been established and the addition of MAG or Nogo66 causes the collapse of growth cones in a dose dependent manner. If NgR27-311, its fragments, and fusion polypeptides including NgR27-311 are active is biologically active then they should inhibit the MAG and Nogo66 mediated growth cone collapse. Growth cone collapse data may be collected by the inspection of photomicrographs of DRG neurons in response to Nogo66 and MAG.
[0071] The L1 polypeptide is a member of the immunoglobulin superfamily of cell adhesion molecules and is expressed in growing axons, glial progenitor cells and Schwann cells throughout life, but has only limited expression in the CNS. L1 interacts with itself and with other extracellular molecules, such as the FGF receptor to promote neurite fasciculation and growth. Expression of L1 is generally associated with a permissive environment for axonal regeneration the, precise region of interest, or fragments of L1 can be derived from the initial clone. For example, Schwann cells that express L1 support peripheral nerve regeneration and axonal growth is observed in the optic nerves from transgenic mice that express L1 in astrocytes but not in wild-type optic nerves. Fibroblasts engineered to express L1 support axonal growth when transplanted into spinal cord. Finally, a soluble form of L1 linked to Fc promoted functional recovery following acute SCI. HU is linked to proteoglycan degrading molecule like chondroitinase ABCI in a fusion protein it is reasonable to expect the fusion protein to promote axonal regeneration in chondroitinase digested regions of a spinal cord injury.
[0072] The neuregulins and their receptors comprise a diverse growth factor and receptor tyrosine kinase system that has been demonstrated to be essential for organogenesis in the CNS, muscle epithelial and other tissues. GGF2 is a soluble isoform of the neuregulin 1 gene. It was initially characterized as a Schwann cell mitogen 32, but subsequent studies have demonstrated direct actions on oligodendrocytes, neurons and other cell types. GGF2 diminishes demyelination and inflammation and enhances remyelination in a mouse model for multiple sclerosis. Based on these results, it is reasonable to expect that a recombinant human GGF2 is a potential treatment for demyelination associated with SCI. With GGF2 is linked to a proteoglycan degrading molecule like chondroitinase ABCI it is reasonable to expect to promote remyelination of axons in chondroitinase-digested regions of a SCI.
TABLE-US-00002 TABLE 2 Examples of non-limiting GGF2 fragments for expression in E. coli for use in compositions and fusion polypeptide compositions. Nucleotide Amino Construct Domain Seq Acid Seq Mol. Wt. GGF2-FL Full length Ml-E422 46 kDa GGF2 GGF2/1 Ig domain + EGF 748-1206 L250-C402 16.8 kDa domain GGF2/2 Ig domain to C- 748-1266 L250-E422 19 kDa terminus GGF2/3 EGF domain 1048-1206 T350-C402 5.7 kDa
[0073] The efficacy of a composition of the present invention, such as a proteoglycan degrading molecule and a molecule that blocks the activity of an axon growth inhibitor either as a mixture [of] or as a fusion protein can be evaluated using a validated rat SCI model at three different levels of SCI severity.
[0074] A proteoglycan degrading molecule like Chondroitinase ABCI is commercially available in small quantities as a naturally derived enzyme (Seikagaku Corporation) and can be made by a recombinant production system to have essentially the same activity as the enzyme purified from Proteus vulgaris. Genomic DNA can be isolated from Proteus vulgaris using DNeasy Tissue kit (Qiagen). PCR primers can be synthesized with an NdeI restriction site at the 5' end and a BamHI site at the 3' end having sequences 5'-CAT ATG GCC ACC AGC MT CCT GCA TTT G-3'(F2) (SEQ ID NO: 95) and 5'-GGA TCC TCA AGG GAG TGG CGA GAG-3'(R) (SEQ ID NO: 96) respectively, to synthesize the mature protein. The 3.0 kb PCR products can be ligated into pCR 2.1 vector (TOPO cloning kit, Invitrogen) and transformed into DH5a competent cells (Invitrogen). Plasmid DNA can be isolated from a number of clones screened by digestion with EcoRI restriction enzyme. The integrity of a gene prepared in this way can be confirmed by repeated DNA sequencing.
[0075] The chondroitinase ABCI sequence can cloned into a PET vector (Novogen) for expression in E. Coli After induction of gene expression with IPTG the bacteria can lysed by sonication with the concomitant extraction of chondroitinase ABCI with Triton X-114/PBS. The inventors discovered that the majority of recombinant chondroitinase ABCI was found in the cytosolic fraction of the bacterial cell lysate that enabled the development of a chondroitinase ABCI purification protocol that yields an enzyme with high activity at high yields. The protocol includes cation-exchange chromatography as a capture step and gel filtration as a polishing step. After these steps chondroitinase ABCI reaches a purity of ˜95%. Anion exchange membrane filtration (Intercept Q, Millipore) can be used for endotoxin and host DNA removal. This step is expected to remove approximately 75% of the endotoxin. Following filtration, chondroitinase ABCI can be dialyzed into volatile buffer, pH 8.0 and lyophilized to dryness. The final product is stable at -70° C. for long term storage. The purified cABCI is a highly basic protein with pI-9.5 as determined by IEF-PAGE analysis of the samples from the crude cell lysate.
[0076] A variety of analytical methods can be used to compare the enzymatic activity of the recombinant version of chondroitinase ABCI to that of a commercially available form of the enzyme (Seikagaku Corporation) purified from Proteus vulgaris. The methods may be adapted to evaluate the activity of fusion proteins including proteoglycan degrading polypeptides like chondroitinase. Specific activity measurements were obtained using an accepted spectrophotometric assay that measures the change in absorbance due to the production of reaction products from the degradation of proteoglycans. The recombinant form of chondroitinase ABCI had approximately 25% higher specific activity than the Seikagaku chondroitinase ABCI. Size exclusion chromatography can be used to compare the hydrodynamic properties the enzymes. The elution profiles for recombinant enzyme was identical to that of the naturally derived enzyme.
[0077] A form of zymography can used to further characterize the enzyme and may be adapted for characterization of the fusion proteins. Polyacrylamide gels can be polymerized in the presence of aggrecan, a substrate for chondroitinase ABCI. Enzyme samples may be resolved on the aggrecan-impregnated gels by electrophoresis in the presence of SDS. The gels can then be subjected to a renaturation step wherein the SDS was extracted and the enzymes allowed to refold. Enzyme refolds and regains activity then digests aggrecan within the gel and the resulting loss of carbohydrate in that region of the gel can be visualized by a carbohydrate-specific stain. A similar loss of carbohydrate in the gel would be expected for active forms of a fusion protein including a proteoglycan degrading polypeptide portion. In the case of recombinant Chondroitinase ABCI, its activity can be visualized as a clear spot in the zymogram. The zymography results were consistent with the spectrophotometric analysis that demonstrates that the recombinant form of chondroitinase ABCI has the same or greater specific activity as the naturally occurring form.
[0078] HPLC methods may be used for detecting the four and six sulphated disaccharides (A4DS and A6DS, respectively) liberated as a result of chondroitinase ABCI digestion of CSPG. The two disaccharides can be effectively resolved by anion exchange chromatography. The HPLC assay has been validated by showing that the quantitation of A4DS and A6DS from chromatograms yields a linear relationship to the amounts injected into the HPLC. Production of A4DS and A6DS from CSPG digestion is directly related to the amount of chondroitinase specific activity as determined by the spectrophotometric assay described above. This assay may be used as a sensitive and accurate means to independently quantitate A4DS and A6DS released by chondroitinase digestion of a variety of substrates and may also be used to determine the activity of chondroitinase polypeptides in a fusion protein.
[0079] Another functional assay that can be performed to characterize proteoglycan polypeptide activity is where dorsal root ganglian (DRG) neurons are plated on aggrecan or aggrecan treated with a proteoglycan like chondroitinase ABCI. It is expected that neurons plated on aggrecan will failed to adhere to the plate and extend axons. In contrast, neurons plated on aggrecan treated with a proteoglycan degrading polypeptide like chondroitinase ABCI in a composition or as part of a fusion polypeptide would be expected to adhere to the surface and extend axons. The extensive axon growth, which is observed for chondroitinase ABCI is believed to be due to the digestion of the carbohydrates on the aggrecan core protein which creates a more permissive substrate for axon growth.
[0080] The rat contusion model of SCI is a clinically relevant model and may be used to evaluate the efficacy of fusion proteins and other compositions of the present invention for promoting axon regeneration. With a contusion SCI, cells are destroyed, hemorrhage ensues and inflammation begins. Destroyed cells are removed by macrophages and a reactive gliosis begins. A cystic cavity is formed and the gliosis matures into a glial scar. Myelin in the area is destroyed and many local neurons that are not destroyed are left in a demyelinated state.
[0081] The forceps compression model of SCI is a contusion model developed and characterized. This model has been validated and results in injuries that are very similar to the more widely used impactor models. The model can involves a forceps compression at vertebral level T9/T10. The forceps compress the cord to a width of 0.9, 1.3 or 1.7 mm for 15 seconds. These three levels of compression allow a severe, mild or moderate injury. This model has been validated using the open field locomotor testing and the Basso, Bresnahan and Beattie (BBB) scoring system. It was also characterized histologically. Behavioral testing and BBB scoring demonstrated that the forceps produce a highly reproducible injury, with recovery similar to that seen with impactor models. These testing and scoring data demonstrate that the forceps compression model is comparable to other contusion models and sufficiently reproducible to be used as an experimental model of SCI and may be used for the evaluation of compositions of the present invention.
[0082] Tissue from forceps compression injured animals can be processed histologically to examine white matter sparing, glial scar and cyst formation. As with other contusion SCI models, a central cyst is formed after injury, with a size that increases with increased injury severity (decrease forceps gap). Around the cyst a glial scar forms that is characterized by astrogliosis (GFAP), macrophage activation and myelin wasting.
[0083] Various aspects of the present invention will be illustrated with reference to the following non-limiting examples.
Example 1
[0084] This example illustrates how a proteoglycan degrading molecule may be administered, studied, and demonstrated to show a functional improvement in animals having a model contusion injury.
[0085] The forceps compression model of SCI is a contusion model developed and characterized. This model has been validated and results in injuries that are very similar to the more widely used impactor models. The model can involves a forceps compression at vertebral level T9/T10. The forceps compress the cord to a width of 0.9, 1.3 or 1.7 mm for 15 seconds. These three levels of compression allow a severe, mild or moderate injury.
[0086] Rats were injured with the forceps compression model at vertebral T9/T10/ At the time of surgery an intrathecal catheter was placed for delivery of chondroitinase. Animals were treated every day for one week and then on alternating days for one week with 0.06 U/dose chondroitinase ABC I (Seikagaku), penicillinase or artificial cerebrospinal fluid (aCSF). Doses and controls were derived from Bradbury et al., 2002. Behavior was assessed using open-field locomotor testing and the BBB scoring system at day 2 and then weekly post injury for ten weeks.
[0087] FIG. 5A shown are mean BBB scores for animals treated with chondroitinase ABC I, penicillinase and aCSF. Rats treated with aCSF or penicillinase recovered to a mean BBB score of about 4. Rats treated with chondroitinase recovered to a mean BBB score of about 8. Multiple animals recovered to scores above 10, indicating supra-spinal input. The chondroitinase scores were significantly different from both control groups by ANOVA and post hoc Tukey.
[0088] Tissue from these animals was processed immunohistochemically for glial fibrillary acidic protein (GFAP) to assess general scar architecture. Tissue was also stained with a Weil stain and a silver degeneration stain to assess myelin and neuron degeneration, respectively. Interestingly no obvious differences in these parameters were noted between experimental and control treated tissues.
[0089] In FIG. 5B the large lesion comprising several segments and the sparing in the ventral cord. Weil staining revealed extensive demyelination in both the treated and untreated animals. The GFAP images (middle set) demonstrate the extent of the scar that is formed following forceps injury. The bottom amino cupric silver degeneration stain demonstrates the vast neural degeneration extending both rostrally and caudally after injury. Again, no obvious differences were noted between chondroitinase treated and control tissues.
[0090] Additional preliminary experiments have been performed at moderate injury levels and significant improvement in open-field locomotor activity was observed with chondroitinase treatment. Animals receiving chondroitinase ABCI recovered to a mean BBB score of 9.1 at ten weeks following injury, compared with 7.1 for the penicillinase controls. The consistency of the data (SEM's of 0.6 and 0.3, respectively) and the region of scores on the BBB scale make this 2 point change not only statistically significant, but also clinically meaningful. A score of 9 indicates plantar placement with weight support or frequent to consistent weight support with dorsal stepping while a score of 7 indicates movement of each joint in hind limb but with no weight support and no consistent sweeping of limbs. An examination of the individual animal scores at ten weeks shows that 6 of 12 animals in the chondroitinase group recovered to scores of 9 or above, while only one of 12 animals in the penicillinase group recovered to a score of 9. One animal from each group was removed from analysis because its score failed to ever rise above 2.5, indicating an injury severity outside the model norms. FIG. 5C and FIG. 5D includes a scatter plot of scores at 10 weeks for each animal in the penicillinase and chondroitinase treatment groups for the moderate injury. Group means are shown below.
[0091] The results show in this well controlled study, that chondroitinase improves open-field locomotor function in Rats that were injured with the forceps compression model at vertebral T9/T10. Animals recovered to mean BBB scores of 9.7 and 9.9 for the penicillinase and chondroitinase groups, respectively. This study demonstrates a significant effect at severe and moderate injury levels, but not mild injury levels. No significant differences were noted between any groups in the mild injury study (1.3 mm forceps). It is unclear if chondroitinase is not effective with mild injury, if chondroitinase effected changes not adequately assayed by the open-field locomotor testing such as stride length, paw placement, sensory or autonomic functions. Preliminary analysis of histology from animals in this study confirmed placement of the catheters and injuries in each animal. Ongoing experiments sacrificing animals at time points after injury with and without chondroitinase treatment will characterize the CSPG content of the scar and the effects of chondroitinase digestion on stub-antigen expression. Future experiments will expand the battery of behavior tests and examine the cord for evidence of regeneration with tract tracing.
[0092] Two acute toxicity studies were conducted in rats. The first was an intravenous (IV) study wherein rats were injected with 0, 0.2, 0.775 or 7.775 mg/kg chondroitinase ABCI. In the second study, intrathecal (IT) catheters were placed over the spinal cords using the same methods employed in the SCI animal studies and 0.06, 0.6 and 6.0 units of chondroitinase ABCI was delivered through the IT catheters. These doses were 1, 10, and 100-fold greater than the estimated local concentrations of chondroitinase ABCI achieved with IT catheters in the SCI experiments. Animals were monitored for pain and distress and body weights were acquired daily. No overt reactions were observed during or immediately after chondroitinase ABCI dosing. No swelling, inflammation, bruising or necrosis was noted at the injection site for the IV experiment. No changes in body temperature were observed in animals treated via the IT route. No alterations in feeding, grooming or vocalizations were noted. Animals were assessed for motor behavior in an open pool. No abnormalities were noted by the animal care staff or behavioral specialists. Animals displayed no signs of joint tenderness or swelling. There were no significant differences in weight change between the treatment groups. The results demonstrate that chondroitinase ABCI treatment is not associated with acute toxicity using IV and IT doses substantially greater than the efficacious IT doses.
Example 2
[0093] This example describes the preparation of Nogo-receptor agonist, L1 neural cell adhesion protein, and GGF2 polypeptide domains which can be used for compositions and fusion proteins of the present invention.
[0094] A soluble portion of the human Nogo receptor spanning amino acids 27 to 311 (NgR27-311) was selected as it has been shown to inhibit the binding of Nogo66, MAG, and MOG to membrane-bound NgR. Primers were designed flanking this region, and RT-PCR was performed using human hippocampal RNA (BD Biosciences). The 1.05 kb region was successfully amplified and purified.
[0095] L1 was prepared through a CHO cell line from the laboratory of Dr. Melitta Schachner that secretes human L1 as a fusion protein with human Fc (L1-Fc). The cells were grown in roller bottles and then L1-Fc was purified from the conditioned media using protein A affinity column chromatography. The purity of L1-Fc was assessed by SDS-PAGE and a single band at the appropriate molecular weight was observed. The biological activity of L1-Fc was confirmed using a neurite outgrowth assay. Tissue culture plates were coated with either poly L-Lysine or L1-Fc and then cerebellar granule cells from postnatal day 10 rats were isolated and placed into culture on the substrates. It was observed that neurons plated on the L1-Fc substrate exhibited a substantial number of long neuritis compared to the polylysine substrate controls. These results demonstrate that the L1-Fc produced is biologically active in promoting neurite outgrowth. This neurite outgrowth assay will be used to assess the biological activity related to the L1 portion of the chondroitinase ABCI fusion protein.
[0096] A CHO cell line that secretes a soluble form of full length GGF2 glycoprotein was obtained from CeNes. Extensive optimization of media and purification methods was completed to obtain essentially pure and biologically active GGF2. A number of analytical methods were developed to characterize the GGF2 including SDS-PAGE, isoelectric focusing, peptide mapping and carbohydrate analysis. For example an SDS-PAGE gel of reduced and non-reduced GGF2; the isolate is essentially free of contaminating proteins and shows the expected molecular weight and monomeric structure. The biological activity of GGF2 was assayed using a primary rat Schwann cell proliferation method and the expected effect was reproducibly obtained with four independent batches of GGF2. Another functional assay was developed that measures the phosphorylation of Akt kinase, a downstream cell signaling component of the erbB receptor pathway. It was observed that there is a dose dependent phosphorylation of Akt kinase
Example 3
[0097] This example illustrates the manipulation of chondroitinase ABCI for construction of chimeras.
[0098] Chondroitinase ABCI was cloned from P. vulgans and expressed in E. coli. Surprisingly repeated attempts to create N-terminal fusion proteins were not successful because the N-Terminal fusion portion was cleaved from chondroitinase ABCI during synthesis.
[0099] However, catalytically active deletion mutants with N-terminal His fusion tags designated ABC I-NΔ2O, ABC I-NΔ4O, and ABCI-NΔ6O were prepared by deleting 20, 40 and 60 amino acids respectively from the N-terminus of the mature ABCI protein. Unlike the full length chondroitinase ABCI, the N-terminal deletion mutants are capable of synthesizing a 6xHis tag as an N-terminal fusion protein. It was also observed that deletion of 80 amino acids from the C-terminal end formed a mutant with proteoglycan degrading activity of chondroitinase ABCI as tested in a zymography assay.
[0100] Fusion proteins with NgR27-311 or L1 with a proteoglycan degrading molecule such as chondroitinase ABCI will utilize mammalian expression and can be performed in CHO cells. The cDNA for chondroitinase ABCI has been cloned in pSECTag vector (Invitrogen) in the proper reading frame. A CHO cell line having secretary chondroitinase ABCI can be developed and the conditioned medium can be tested for catalytic activity by zymography assay to confirm that chondroitinase ABCI expressed in mammalian cells is functional. A mammalian cell codon optimized version of chondroitinase ABCI can be synthesized by methods known in the art for use for CHO cell expression.
[0101] GGF2 is a spliced variant of the NRG1 gene expressed in brain and spinal cord of adult humans. It is a glycosylated protein of molecular mass between 66-90 kDa. The inventors have discovered that recombinant GGF2 expressed in CHO cells is highly glycosylated and promotes Schwann cell proliferation in vitro and further that an EGF-like domain of NRG1 expressed in E. coli is fully functional in promoting myocyte proliferation and survival.
[0102] The inventors have expressed fragments of GGF2 in E. coli as described. The specific GGF2 domain responsible for Schwann cell proliferation and thereby remyelination can be determined. If the Ig and EGF domains together or separately show biological activity in vitro, then they can be used to form chimeric fusion proteins.
[0103] Cloning and expression of NgR27-311 in CHO cells. An NgR fragment corresponding to residues 1-359 was isolated by RT-PCR from human hippocampus poly K RNA (BD Biosciences) and its structure can be confirmed by DNA sequencing. The gene fragment corresponding to residues 27-311 can be cloned from the larger fragment and then subcloned in pSECTag vector (Invitrogen) in the proper reading frame to express the fragment as a secretary protein in CHO cells. The plasmid DNA containing the NgR27-311 gene can be transfected in CHO cells and the cell line producing NgR27-311 can be selected under the selection pressure of hygromycine B. An NgR27-311-ABCI chimera expression plasmid can be constructed for expression in a CHO cell expression system using methods known in the art.
Example 4
[0104] This example illustrates methods which may be used to purify and isolate expressed chondroitinase ABCI, and GGF2 domains expressed in E. coli. These method may be applied to purification of chimeric fusion proteins of the present invention.
[0105] An efficient E. coli recombinant expression system and a purification process for chondroitinase ABCI have been developed by the inventors that could be applied for the purification of chondroitinase ABCI chimeric derivatives.
[0106] For example, the expression of various GGF2 domains in the chondroitinase ABCI expression host, E. coli has been performed and the following peptides have been tested: aa250-402, aa250-422 and aa350-402. These expressed peptides were found in the soluble fraction of the E. coli lysates. It is reasonable to expect that final chimeric products of these peptides with chondroitinase ABCI in E. coli will also be soluble. In addition, it is expected that the charge characteristics of the chimeric products when compared to the recombinant chondroitinase ABCI will be similar. Thus, the theoretical isoelectric point (p1) values for the GGF2 peptides are 9.3 for aa250-402, 9.18 for aa250-422 and 7.55 for aa350-402. Fusing the first two peptides with chondroitinase ABCI is expected to result in chimeric proteins with p1 values still above 9. In this case SP chromatography is expected to perform well as the capturing step. The smallest GGF2 peptide, aa 350-402, will reduce the p1 of the final chondroitinase ABCI chimera to approximately 8.4. This change may require optimization of the capture step conditions.
[0107] The chimeric products of chondroitinase ABCI with NgR27-311 and L1 peptides can be expressed in a CHO cell system. Prior to purification of the expressed chimeric proteins, the growth conditions for the cell line producing either the NgR27-311 or L1 chimeric proteins will be optimized in serum-free medium. A detailed media optimization study can be performed to determine the highest production conditions. Scale-up volume can be decided based on the rate of production of the chimeric proteins (pg/cell/day). Conditioned media from the various chimera-producing cell lines can be collected and subjected to tangential flow filtration. Ion-exchange chromatography can be used for capturing the secreted proteins from conditioned media, gel filtration chromatography can be used as a polishing purification step and then anion-exchange membrane filtration can be used for endotoxin and DNA removal. At each step the efficiency of the purification will be analyzed by SDS-PAGE, and spectrophotometric quantitation of protein concentration.
Example 5
[0108] This prophetic example describes in vitro assessment of chimera biological activity: Each chimera can be assayed for chondroitinase enzymatic activity and the specific biological activity of each fusion partner.
[0109] The first step in analysis may employ conventional protein biochemical methodologies to confirm the fidelity of gene expression. These include SDSPAGE, lEE, mass spectrometry and size exclusion chromatography.
[0110] Chondroitinase chimera specific activity can be determined using a standard and uniformly accepted spectrophotometric assay. The production of reaction products from the catalytic activity of a chondroitinase chimeric polypeptide can be determined by a measurement of the absorbance of the product at a wavelength of 232 nm. A typical reaction mixture consists of 120 microliters of reaction mixture (40 mM Tris, pH 8.0, 40 mM NaAcetate, 0.002% casein) combined with substrate (5 microliters of 50 mM chondroitin C) and 1.5 microliters of chondroitinase ABCI chimeric fusion polypeptide or test sample. The change in absorbance units is the initial rate which can be converted to a unit activity measurement.
[0111] Disaccharide HPLC assay: The catalytic activity of chondroitinase chimeric polypeptide on a CSPG substrate is expected to releases two species of sulphated disaccharides including α-Δ AUA-[1→3]-GaINAc-4S (Δ 4DS) and α-Δ AUA-[1→3]-GaINAc-6S (Δ 6DS). These species can be resolved by HPLC and the quantitation from the resulting chromatograms is a sensitive and accurate measure of chondroitinase activity. To perform the assay, samples from chondroitinase digestion reactions carried out with wild-type chondroitinase and chondroitinase fusion proteins can be clarified by centrifugation and then subjected to a validated anion exchange HPLC method as follows. A Dionex CarboPac PA-10 analytical column (4×250 mm) fitted with a Dionex CarboPac PA-10 (4×50 mm) guard column can be used with a mobile phase consisting of a gradient of water at pH 3.5 (Buffer A) and 2M NaCl, at pH 3.5 (Buffer B). Detection may be set at a wavelength of 232 nm. A flow rate of 1 mL/minute and a 45 minute continuous gradient of 100% A to 100% B affords acceptable resolution of Δ4DS and Δ 6DS. Standard curves can be generated using known amounts of Δ4DS and Δ6DS.
[0112] Zymography allows resolution of proteins by molecular weight with a concomitant assessment of chondroitinase activity. A 10% polyacrylamide gel may be polymerized in the presence of 85 pg/ml of aggrecan. Samples can be boiled in an SDS-loading buffer and then the analytes can be resolved by electrophoresis. After separation the gel can be incubated for 1 hour at room temperature in 2.5% Triton X100 then 16 hours at 37° C. in fresh 2.5% Triton X-100. During these incubations, the SOS can be extracted from the gel and the chondroitinase refolds and digests the aggrecan in its immediate vicinity. After the refolding process the gel can be stained for carbohydrate: The gel can be first incubated in 0.2% Cetylpyridinium for 90 minutes at room temperature and then transferred into 0.2% Toludine Blue in 49:50:1 H20, Ethanol, and Acetic Acid for 30 minutes. The gel can then be fully destained. Following destaining the gel can be incubated overnight in a 50 microgram/ml solution of Stains-All (Sigma) in 50% ethanol. Chondroitinase activity can be detected as a clear spot in the gel that is coincident with the molecular weight of the enzyme. The size of the clearing has been shown to be almost linearly related to Unit activity.
[0113] The NgR27-311-chondroitinase chimera can be assessed for the activity of the decoy Nogo receptor. The assay can be used to measures the collapse of growth cones: Dorsal root ganglia (DRG) are dissected from postnatal day 1 (P1) Sprague Dawley rat pups and dissociated in 200 U/ml collagenase I (Worthington) and 2.5 U/ml dispase (Boehringer/Roche) 2 times for 30 min. at 37° C. Enzymes are removed and DNAse (0.5 mg/ml) can be added to the ganglia. Trituration may be done with a pipette tip attached to a 1000 μl Pipetman. The resulting cell suspension can be filtered through a 40 micron cell filter and centrifuged at 70×g for 5 min. Cells can be re-suspended in DMEM/10% FBS and pre-plated for 2 hours on a non-coated tissue culture plate (100 mm diameter). Non-adherent neurons are removed and plated at 10,000 cells/well in a Poly-lysine/laminin-coated 24 well plate in serum-free Neurobasal/B27 with 50 ng/ml NGF. After 20 to 24 hours, MAG or Nogo66 can be added at varying concentrations for 1 hour at 37° C. to induce growth cone collapse. NgR27-311-chondroitinase chimera can be added at various concentrations to compete with MAG and Nogo66 and thus protect the neurons from growth cone collapse. Cultures may be fixed by adding an equal volume of pre-warmed 8% paraformaldehyde/0.6 M sucrose to the medium for 20 min. while the cells are kept on a 37° C. hot plate. Growth cones may be labeled with AlexaS68 phalloidin (Molecular Probes). Briefly, cells can be permeabilized with 0.1% Triton-X 100 for 5 min. at RT, blocked for 20 min. in 1% BSA in PBS and incubated in phalloidin, diluted 1:40 in 1% BSA, in PBS for 20 mm at RT. Cells can be washed in PBS and mounted in Fluorescent Mounting Medium (DAKO). The percentage of collapsed growth cones is determined by analyzing a minimum of 100 growth cones per well under a 40× objective.
[0114] The biological activity L1 fusion proteins may be determined using a standardized neurite outgrowth assay. After etching a 25 mm circle in the center of a tissue culture-treated 35 mm dish, 1 ml of 10 μg/ml poly-lysine can be added to each etched circle and incubated at 37° C. for 60 minutes. The poly-lysine provides a negative control for neurite outgrowth. Etched circles may be washed 2 times with Hank's balanced salt solution plus calcium and magnesium (HBSS++) after the last rinse; 1.2 μl of L1-Fc (0.6 μM, 0.3 μM, 0.15 μM, 0.075 μM) is spotted onto the petri dishes to serve as a positive control and various concentrations of the L1-chondroitinase fusion protein can be applied to the plates as test samples. The plates can be incubated for 1 hour at room temperature. The spots are lightly aspirated, so as not to dry them out, and immediately washed 2 times with 1 ml HBSS++ and then 1 ml of 1% BSA in PBS is added to each etched circle. After about 15 minutes at room temperature the etched circles can be washed 2 times with HBSS++, and once with bioassay medium (NeuralBasal (Gibco)+B27 supplements (Gibco)+L-glutamine+L-glutamic acid, penicillin and streptomycin (100 U/ml)+1% fetal bovine serum which will remain on the petri dishes until the time of the assay. To provide neurons to plate on the substrates Cerebellar granule cells from postnatal day (PND) 9 or 10 are harvested. The brain is removed from the cranium of 1 pup, and the cerebellum separated from the rest of the tissue. Meninges is removed, and the cerebellum is placed in ice cold HBSS++. The tissue is minced and trypsinized using 0.25% trypsin for 15 minutes at 37° C. Trypsin action is inhibited by adding 0.5 mg/ml soybean trypsin inhibitor (Gibco). The tissue is rinsed with HBSS++ and triturated using a flame narrowed Pasteur pipette coated with EBS. Dissociated cells are pelleted at 500×g for 3 minutes, the supernatant is decanted and the cells are resuspended in 2 ml of growth medium. After lightly triturating, the cell suspension is carefully layered on top of a 3.5% BSA (in PBS) cushion, and spun at 500×g for 3 minutes. Supernatant is aspirated and the pellet is resuspended in 2 ml growth media. A cell count is performed and the cells are diluted to a final working concentration of 1.5×105 cell/ml. A 300 μl aliquot of diluted cells is added to the center of each plate, resulting in the even distribution of 6×104 cells across the entire surface area of the etched circle. The cells are allowed to grow for 16 hours at 37° C. in a humidified environment supplemented with 5% CO2. The next day, plates are removed from the 37° C. incubator, cells are fixed with 4% paraformaldehyde and the outgrowth of neurites is recorded by photomicroscopy.
[0115] Biological activity assays for GGF2 may be performed using Schwann cell proliferation. Sciatic nerves are dissected from three day old Sprague Dawley rat pups and dissociated in L-15 medium (Invitrogen) containing 0.25% trypsin and 0.03% collagenase type I (SIGMA) for 15 minutes at 37° C. Nerves are centrifuged at 400×g for 5 minutes and dissociation medium is replaced by DMEM/10% FBS. Nerves are triturated using a 10 ml syringe with a 21 g needle and subsequently a 23 g needle. The cell suspension is filtered through a 40 μm mesh placed over the opening of a 50 ml conical tube. Cells can be centrifuged at 400×g for 5 min, plated in a poly-D-lysine (PDL) coated T-75 flask at a density of approximately 5 million cells in 15 ml of DMEM/10% FBS/PenStrep and incubated in a 37° C. incubator regulated with 10% CO2. After a 24 hour incubation the cells can be washed twice with DMEM/10% FBS and re-fed with fibroblast inhibition medium consisting of DMEM/10% FBS and 10 μl/ml of 1 mM cytosine arabinoside (Ara-C). After an incubation time of 2 to 3 days the fibroblast inhibition medium is replaced with Schwann cell growth medium (DMEM/10% FBS/150 ng/ml of GGF2/5 μM forskolin). Cells can be expanded and aliquots of 2×106 cells/ml are frozen in DMEM/10% DMSO, 54% FBS in liquid nitrogen. At the time of use, Schwann cells are thawed and plated at a density of about 16,000 cells/well in a PDL-coated 96 well tissue culture plate in DMEM/5% FBS. After about 24 hours GGF2 is added in serial dilution ranging from 100 ng/ml to 0.78 ng/ml containing 5 μM forskolin to establish a standard curve. Samples of GGF2 fusion proteins can also be added in serial dilution together with 5 μM forskolin. BrdU is added at 10 μM. Cells are incubated for 48 hours in a 10% CO2 incubator. A BrdU ELISA kit from Roche Applied Science can be used (Cat. No. 1 647 229) to detect Schwann cell proliferation. Medium is poured out of the plate and plate is tapped on tissue paper. A 200 μl/well aliquot of Fix/Denat is added and incubated for 30 min at 15-25° C. A 100 μl/well aliquot of anti-BrdU-POD working solution (lyophilized antibody is dissolved in 1.1 ml double distilled water and diluted 1:100 with antibody dilution solution) can be added and incubated for 90 min at room temperature. Wells may be washed 3 times with 200-300 μl/well of washing solution. A 100 μl aliquot of substrate solution is added and incubated for 15 minutes or until color development is sufficient for photometric detection. The Plates are read on a SpectraMax plate reader at 450 nm either before addition of stop solution at 370 nm or after addition of 50 ml 1N sulfuric acid to each well.
[0116] Akt [pS473] ELISA: C6 glioma cells, obtained from ATCC, are grown in DMEM/10% FBS in a T-75 flask to confluence. After trypsinization, cells are plated in a 24 well plate at a density of 500,000 cells/well in 0.5 ml of medium. One day after plating the cells are treated with GGF2 (batch: Glu from Lonza Biologics) at serial dilutions ranging from 0.78 to 100 ng/ml for 30 minutes at 37° C. to establish a standard curve. As a negative control, wortmannin, a PI3-kinase inhibitor, is added to cells at 10 nnM for 30 minutes before the addition of GGF2. Samples of GGF2 fusion proteins will also be added in serial dilution. The cells are washed with PBS and then extracted with 100 μl of cell extraction buffer (10 mM Tris, pH 7.4, 100 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1 mM NaF, 20 mM Na4P2O7, 2 mM Na3VO4, 1% Triton X-100, 10% glycerol, 0.1% SDS, 0.5% deoxycholate, 1 mM PMSF, protease inhibitor cocktail (Pierce)). Cell extracts are kept at -80° C. until further use. An ELISA kit from Biosource (Cat. #KHOO1 II) can be used to measure phosphorylated Akt kinase levels. Briefly, 100 μl of a 1:50 dilution of samples and a serial dilution of Akt [pS473] standards are loaded into wells pre-coated with anti-Akt antibody for 2 h at room temperature (RT) or overnight at 4° C. Wells are washed and anti-Akt [pS473] antibody is added and incubated for 1 hour. After washing, HRP-conjugated anti-rabbit antiserum is added to wells and incubated for 30 min. After washing, TMB chromogen is added to wells and incubated for 30 min at RT before the reaction is stopped with stop solution. The plate is read on a SpectraMax plate reader at 450 nm. The optical densities are plotted against the Akt [pS473] standards and the concentration of pAkt in the C6 samples are deduced from the standard curve.
Example 6
[0117] This example illustrates construction of a fusion protein of chondroitinase polypeptide and a TAT cellular transduction peptide.
[0118] The gene sequence encoding a chondroitinase enzyme can be functionally linked to the protein transduction domain from HIV called the TAT Peptide (SEQ ID NO: 61). The resultant chimeric gene TAT-chondroitinase ABCI fusion DNA construct is shown in FIG. 1. It was observed that during bacterial expression of this construct, the TAT peptide was removed from the chondroitinase enzyme at some processing point during the bacterial growth. The removal of n-terminal linked peptides was also observed during expression of an n-terminal histidine-tagged chondroitinase ABCI enzyme.
[0119] Deletion mutants of the chondroitinase ABCI enzyme were generated where the mutant was missing a certain number of amino acids from the n-terminal portion of the polypeptide but maintained it proteoglycan degrading activity. It was observed that these n-terminal deletion maintained a histidine-tag that was attached to the n-terminus. It is expected that various TAT-deletion mutant chondroitinase ABCI fusion DNA construct can be expressed without removal of the TAT polypeptide during expression. For example, the TAT peptide may be fused at the N-terminus of deletion mutants like ABCI-NΔ20 or ABCI-NΔ60 deletion mutant. Without wishing to be bound by theory, the inventors believe that the native proteoglycan degrading enzyme contains a signal sequence that is attached to the n-terminus. This signal sequence is removed during the natural production in bacteria and in production of the cloned enzyme in E. coli. It is thought that some signal within the n-terminal amino acids instructs the bacteria to remove anything attached to this end.
[0120] A DNA construct with the TAT-peptide attached to the N-terminus of one of the chondroitinase deletion mutants can be made and Western blot and protein gel showing this expressed protein and activity.
Example 7
[0121] This example illustrates the diffusion of molecules into cells and tissue using a proteoglycan degrading composition.
[0122] A brain from an adult Sprague Dawley rat was removed from the skull and quartered into Right frontal, left frontal, right rear and left rear sections, corresponding roughly to the frontal (front) and occipito-parietal (rear) lobes. (A) Right frontal quarter was placed in artificial cerebrospinal fluid (Catalog #59-7316; Harvard Apparatus, Holliston, Mass.) containing the beta-galactosidase enzyme (Catalog #, G 5160; Sigma, St. Louis, Mo.) and chondroitinase ABC I at 0.5 U/ml (Catalog# C 3667; Sigma, St. Louis, Mo.) for 2 hours at 37° C. Brain quarter was rinsed several times in phosphate buffered saline (PBS) and then processed with a Beta-Gal staining kit (Catalog # Gal-S; Sigma, St. Louis, Mo.). The substrate-enzyme reaction (blue product) was allowed to develop for 1 hour, and the brain was rinsed several times in PBS and slabs from the middle of each brain block cut using parallel straight razors. (B) Left frontal quarter was placed in artificial cerebrospinal fluid (Catalog #59-7316; Harvard Apparatus, Holliston, Mass.) containing the beta-galactosidase enzyme (Catalog #, G 5160; Sigma, St. Louis, Mo.) for 2 hours at 37° C. Brain quarter was rinsed several times in phosphate buffered saline (PBS) and then processed with a Beta-Gal staining kit (Catalog # Gal-S; Sigma, St. Louis, Mo.). The substrate-enzyme reaction (blue product) was allowed to develop for 1 hour, and the brain was rinsed several times in PBS and slabs from the middle of each brain block cut using parallel straight razors. (C) Right frontal quarter was placed in artificial cerebrospinal fluid (Catalog #59-7316; Harvard Apparatus, Holliston, Mass.) containing the beta-galactosidase enzyme (Catalog #, G 5160; Sigma, St. Louis, Mo.) and chondroitinase ABC I at 0.5 U/mi (Catalog# C 3667; Sigma, St. Louis, Mo.) for 2 hours at 37° C. Brain quarter was rinsed several times in phosphate buffered saline (PBS) and then processed with a Beta-Gal staining kit (Catalog # Gal-S; Sigma, St. Louis, Mo.). The substrate-enzyme reaction (blue product) was allowed to develop for 1 hour, and the brain was rinsed several times in PBS and slabs from the middle of each brain block cut using parallel straight razors. (D) Left rear quarter was placed in artificial cerebrospinal fluid (Catalog #59-7316; Harvard Apparatus, Holliston, Mass.) containing the beta-galactosidase enzyme (Catalog #, G 5160; Sigma, St. Louis, Mo.) for 2 hours at 37° C. Brain quarter was rinsed several times in phosphate buffered saline (PBS) and then processed with a Beta-Gal staining kit (Catalog # Gal-S; Sigma, St. Louis, Mo.). The substrate-enzyme reaction (blue product) was allowed to develop for 1 hour, and the brain was rinsed several times in PBS and slabs from the middle of each brain block cut using parallel straight razors. Images of brains were acquired on a scanner and are shown in FIG. 2(I)(A-D).
[0123] Adult rat brain hemispheres were soaked in buffer alone or containing 33 U/ml Chondroitinase ABCI (Acorda) for 2 hours at 37 degrees C. Hemispheres were rinsed and immediately placed in Eosin Y (Sigma) or a saturated solution of Congo Red (Sigma) in 70% ethanol. Slabs of tissue were cut and images were acquired on a scanner. See FIG. 2(II) Eosin and FIG. 2(III) Congo red.
[0124] FIG. 2(III) saturated solution of Congo Red demonstrates greater penetration through the cortex of a Chondroitinase treated brain hemisphere as compared to untreated brain. Congo Red is a negatively charged dye of 697 kDA. FIG. 2(II) Eosin Y penetration through the cortex of a Chondroitinase treated brain hemisphere looks slightly more diffuse, but penetration does not seem to be any deeper as compared to untreated brain. Eosin Y is zwitterionic, having an overall negative charged at the low pH it was used at, and it is 692 kDa. FIG. 2(I) Lobes from adult rat were incubated in beta-galactosidease alone (B&D), or with the addition of Chondroitinase ABCI (A, 0.5 U/ml or C, 0.005 U/m).
[0125] The results showed that chondroitinase treated tissue affected penetration of beta-galactosidase into CNS tissue. Chondroitinse had dramatic effects on the rate of Congo Red dye penetration but apparently not Eosin.
Example 8
[0126] This example illustrates a Chondroitinase ABCI Assay Protocol which may be modified to measure the activity of Chondroitinase ABCI deletion mutant-fusion proteins or the activity of other proteoglycan degrading polypeptide-fusion proteins of the present invention.
[0127] The production of reaction products from the catalytic activity of a proteoglycan degrading molecule or fusion protein is determined by a measurement of the absorbance of the product at a wavelength of 232 nm. A typical reaction mixture consists of 120 μl of reaction mixture (40 mM Tris, pH 8.0, 40 mM NaAcetate, 0.002% casein) combined with a substrate (5 IA of 50 mM chondroitin C) and 1.5 μl of chondroitinase ABCI or a fusion protein of a deletion mutant of chondroitinase ABCI-TAT. Seikagaku's cABCI reference: frozen at -20° C. in 5 ml aliquots, use 1 ml per reaction, 50 mM chondroitin C (MW 521) in water: frozen at -20° C. in 100 ml aliquots. Reaction mixture aliquots of about 120 μl can be prepared at 37° C. for 3 min or longer. A wavelength of 232 nm, is used with the spectrometer.
[0128] Knowing the extinction coefficient for the reaction product, measuring the change in the absorbance of the reaction product at 232 nm reading over time upon addition of a known amount of the Chondroitinase ABCI or other proteoglycan degrading polypeptide-fusion proteins to the 120 μl reaction mixture with 0.002% casein and chondroitin substrate added, the specific activity in μmol/min/mg of the proteoglycan degrading fusion protein can be determined. Seikagaku Chondroitinase ABCI has a specific activity under these assay conditions of about 450 μmole/min/mg.
[0129] Although the present invention has been described in considerable detail with reference to certain preferred embodiments thereof, other versions are possible. Therefore the spirit and scope of the appended claims should not be limited to the description and the preferred versions contain within this specification.
Sequence CWU
1
1
9611021PRTUnknownProteus vulgaris Chondroitinase ABC I protein with
gwra and dalni sequences 1Met Pro Ile Phe Arg Phe Thr Ala Leu Ala Met Thr
Leu Gly Leu Leu 1 5 10
15 Ser Ala Pro Tyr Asn Ala Met Ala Ala Thr Ser Asn Pro Ala Phe Asp
20 25 30 Pro Lys Asn
Leu Met Gln Ser Glu Ile Tyr His Phe Ala Gln Asn Asn 35
40 45 Pro Leu Ala Asp Phe Ser Ser Asp
Lys Asn Ser Ile Leu Thr Leu Ser 50 55
60 Asp Lys Arg Ser Ile Met Gly Asn Gln Ser Leu Leu Trp
Lys Trp Lys 65 70 75
80 Gly Gly Ser Ser Phe Thr Leu His Lys Lys Leu Ile Val Pro Thr Asp
85 90 95 Lys Glu Ala Ser
Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser 100
105 110 Phe Trp Leu Tyr Asn Glu Lys Pro Ile
Asp Gly Tyr Leu Thr Ile Asp 115 120
125 Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala Gln Ala Gly
Phe Lys 130 135 140
Val Lys Leu Asp Phe Thr Gly Trp Arg Ala Val Gly Val Ser Leu Asn 145
150 155 160 Asn Asp Leu Glu Asn
Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser 165
170 175 Ser Asp Gly Thr Gln Asp Ser Ile Gly Arg
Ser Leu Gly Ala Lys Val 180 185
190 Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn Val Ser Gln Gly Glu
Ile 195 200 205 Tyr
Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp 210
215 220 Ser Asp Tyr Gln Val Lys
Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe 225 230
235 240 His Asn Val Lys Pro Gln Leu Pro Val Thr Pro
Glu Asn Leu Ala Ala 245 250
255 Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu Phe Val Gly Gly Glu
260 265 270 Lys Glu
Thr Asn Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser 275
280 285 Asp Phe Asp Ala Leu Asn Ile
His Thr Leu Ala Asn Gly Gly Thr Gln 290 295
300 Gly Arg His Leu Ile Thr Asp Lys Gln Ile Ile Ile
Tyr Gln Pro Glu 305 310 315
320 Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp Asn Tyr Val Ile Leu
325 330 335 Gly Asn Tyr
Thr Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val Leu 340
345 350 Glu Lys Asp Pro Thr Gln Lys Ala
Gln Leu Lys Gln Met Tyr Leu Leu 355 360
365 Met Thr Lys His Leu Leu Asp Gln Gly Phe Val Lys Gly
Ser Ala Leu 370 375 380
Val Thr Thr His His Trp Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser 385
390 395 400 Thr Leu Leu Met
Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln 405
410 415 Val Tyr Asp Ser Leu Leu Trp Tyr Ser
Arg Glu Phe Lys Ser Ser Phe 420 425
430 Asp Met Lys Val Ser Ala Asp Ser Ser Asp Leu Asp Tyr Phe
Asn Thr 435 440 445
Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln 450
455 460 Lys Arg Ile Asn Leu
Val Asn Thr Phe Ser His Tyr Ile Thr Gly Ala 465 470
475 480 Leu Thr Gln Val Pro Pro Gly Gly Lys Asp
Gly Leu Arg Pro Asp Gly 485 490
495 Thr Ala Trp Arg His Glu Gly Asn Tyr Pro Gly Tyr Ser Phe Pro
Ala 500 505 510 Phe
Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe 515
520 525 Ser Val Gly Glu Ser Gly
Trp Asn Asn Leu Lys Lys Ala Met Val Ser 530 535
540 Ala Trp Ile Tyr Ser Asn Pro Glu Val Gly Leu
Pro Leu Ala Gly Arg 545 550 555
560 His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val Ala Gln Gly Tyr Tyr
565 570 575 Trp Leu
Ala Met Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser 580
585 590 Ile Tyr Leu Ala Ile Ser Asp
Lys Thr Gln Asn Glu Ser Thr Ala Ile 595 600
605 Phe Gly Glu Thr Ile Thr Pro Ala Ser Leu Pro Gln
Gly Phe Tyr Ala 610 615 620
Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met Val 625
630 635 640 Thr Leu Lys
Ala Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn 645
650 655 Lys Asp Asn Arg Tyr Gly Arg Tyr
Gln Ser His Gly Val Ala Gln Ile 660 665
670 Val Ser Asn Gly Ser Gln Leu Ser Gln Gly Tyr Gln Gln
Glu Gly Trp 675 680 685
Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys 690
695 700 Asp Leu Asp Ser
Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg 705 710
715 720 Gly Phe Ser Gly Thr Ser Ser Leu Glu
Gly Gln Tyr Gly Met Met Ala 725 730
735 Phe Asn Leu Ile Tyr Pro Ala Asn Leu Glu Arg Phe Asp Pro
Asn Phe 740 745 750
Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile
755 760 765 Gly Ser Asn Ile
Asn Ser Ser Asp Lys Asn Lys Asn Val Glu Thr Thr 770
775 780 Leu Phe Gln His Ala Ile Thr Pro
Thr Leu Asn Thr Leu Trp Ile Asn 785 790
795 800 Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln Thr Thr
Leu Gln Gln Gly 805 810
815 Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala
820 825 830 Glu Lys Val
Asn Val Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys 835
840 845 Asn Arg Gln Pro Thr Glu Gly Asn
Phe Ser Ser Ala Trp Ile Asp His 850 855
860 Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr Met Val
Phe Leu Asp 865 870 875
880 Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys Phe Arg Glu Asn
885 890 895 Asn Gly Leu Tyr
Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile 900
905 910 Leu Asp Lys Leu Ser Asn Val Thr Gly
Tyr Ala Phe Tyr Gln Pro Ala 915 920
925 Ser Ile Glu Asp Lys Trp Ile Lys Lys Val Asn Lys Pro Ala
Ile Val 930 935 940
Met Thr His Arg Gln Lys Asp Thr Leu Ile Val Ser Ala Val Thr Pro 945
950 955 960 Asp Leu Asn Met Thr
Arg Gln Lys Ala Ala Thr Pro Val Thr Ile Asn 965
970 975 Val Thr Ile Asn Gly Lys Trp Gln Ser Ala
Asp Lys Asn Ser Glu Val 980 985
990 Lys Tyr Gln Val Ser Gly Asp Asn Thr Glu Leu Thr Phe Thr
Ser Tyr 995 1000 1005
Phe Gly Ile Pro Gln Glu Ile Lys Leu Ser Pro Leu Pro 1010
1015 1020 286PRTUnknownHuman immunodeficiency
virus HIV-1 TAT protein 2Met Glu Pro Val Asp Pro Arg Leu Glu Pro Trp Lys
His Pro Gly Ser 1 5 10
15 Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Lys Lys Cys Cys Phe
20 25 30 His Cys Gln
Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 35
40 45 Arg Lys Lys Arg Arg Gln Arg Arg
Arg Pro Pro Gln Gly Ser Gln Thr 50 55
60 His Gln Val Ser Leu Ser Lys Gln Pro Thr Ser Gln Ser
Arg Gly Asp 65 70 75
80 Pro Thr Gly Pro Lys Glu 85
311PRTArtificialSynthetic peptide, amino acid residues 47-57 of
(HIV-1) Tat protein, (Vives, et al.) 3Tyr Gly Arg Lys Lys Arg Arg Gln Arg
Arg Arg 1 5 10
4285PRTArtificialSynthetic Peptide derived from Nogo-66 receptor
antagonist Locus (Q9BZR6) 4Cys Pro Gly Ala Cys Val Cys Tyr Asn Glu Pro
Lys Val Thr Thr Ser 1 5 10
15 Cys Pro Gln Gln Gly Leu Gln Ala Val Pro Val Gly Ile Pro Ala Ala
20 25 30 Ser Gln
Arg Ile Phe Leu His Gly Asn Arg Ile Ser His Val Pro Ala 35
40 45 Ala Ser Phe Arg Ala Cys Arg
Asn Leu Thr Ile Leu Trp Leu His Ser 50 55
60 Asn Val Leu Ala Arg Ile Asp Ala Ala Ala Phe Thr
Gly Leu Ala Leu 65 70 75
80 Leu Glu Gln Leu Asp Leu Ser Asp Asn Ala Gln Leu Arg Ser Val Asp
85 90 95 Pro Ala Thr
Phe His Gly Leu Gly Arg Leu His Thr Leu His Leu Asp 100
105 110 Arg Cys Gly Leu Gln Glu Leu Gly
Pro Gly Leu Phe Arg Gly Leu Ala 115 120
125 Ala Leu Gln Tyr Leu Tyr Leu Gln Asp Asn Ala Leu Gln
Ala Leu Pro 130 135 140
Asp Asp Thr Phe Arg Asp Leu Gly Asn Leu Thr His Leu Phe Leu His 145
150 155 160 Gly Asn Arg Ile
Ser Ser Val Pro Glu Arg Ala Phe Arg Gly Leu His 165
170 175 Ser Leu Asp Arg Leu Leu Leu His Gln
Asn Arg Val Ala His Val His 180 185
190 Pro His Ala Phe Arg Asp Leu Gly Arg Leu Met Thr Leu Tyr
Leu Phe 195 200 205
Ala Asn Asn Leu Ser Ala Leu Pro Thr Glu Ala Leu Ala Pro Leu Arg 210
215 220 Ala Leu Gln Tyr Leu
Arg Leu Asn Asp Asn Pro Trp Val Cys Asp Cys 225 230
235 240 Arg Ala Arg Pro Leu Trp Ala Trp Leu Gln
Lys Phe Arg Gly Ser Ser 245 250
255 Ser Glu Val Pro Cys Ser Leu Pro Gln Arg Leu Ala Gly Arg Asp
Leu 260 265 270 Lys
Arg Leu Ala Ala Asn Asp Leu Gln Gly Cys Ala Val 275
280 285 51257PRTUnknownHomo sapiens L1 protein 5Met Val
Val Ala Leu Arg Tyr Val Trp Pro Leu Leu Leu Cys Ser Pro 1 5
10 15 Cys Leu Leu Ile Gln Ile Pro
Glu Glu Tyr Glu Gly His His Val Met 20 25
30 Glu Pro Pro Val Ile Thr Glu Gln Ser Pro Arg Arg
Leu Val Val Phe 35 40 45
Pro Thr Asp Asp Ile Ser Leu Lys Cys Glu Ala Ser Gly Lys Pro Glu
50 55 60 Val Gln Phe
Arg Trp Thr Arg Asp Gly Val His Phe Lys Pro Lys Glu 65
70 75 80 Glu Leu Gly Val Thr Val Tyr
Gln Ser Pro His Ser Gly Ser Phe Thr 85
90 95 Ile Thr Gly Asn Asn Ser Asn Phe Ala Gln Arg
Phe Gln Gly Ile Tyr 100 105
110 Arg Cys Phe Ala Ser Asn Lys Leu Gly Thr Ala Met Ser His Glu
Ile 115 120 125 Arg
Leu Met Ala Glu Gly Ala Pro Lys Trp Pro Lys Glu Thr Val Lys 130
135 140 Pro Val Glu Val Glu Glu
Gly Glu Ser Val Val Leu Pro Cys Asn Pro 145 150
155 160 Pro Pro Ser Ala Glu Pro Leu Arg Ile Tyr Trp
Met Asn Ser Lys Ile 165 170
175 Leu His Ile Lys Gln Asp Glu Arg Val Thr Met Gly Gln Asn Gly Asn
180 185 190 Leu Tyr
Phe Ala Asn Val Leu Thr Ser Asp Asn His Ser Asp Tyr Ile 195
200 205 Cys His Ala His Phe Pro Gly
Thr Arg Thr Ile Ile Gln Lys Glu Pro 210 215
220 Ile Asp Leu Arg Val Lys Ala Thr Asn Ser Met Ile
Asp Arg Lys Pro 225 230 235
240 Arg Leu Leu Phe Pro Thr Asn Ser Ser Ser His Leu Val Ala Leu Gln
245 250 255 Gly Gln Pro
Leu Val Leu Glu Cys Ile Ala Glu Gly Phe Pro Thr Pro 260
265 270 Thr Ile Lys Trp Leu Arg Pro Ser
Gly Pro Met Pro Ala Asp Arg Val 275 280
285 Thr Tyr Gln Asn His Asn Lys Thr Leu Gln Leu Leu Lys
Val Gly Glu 290 295 300
Glu Asp Asp Gly Glu Tyr Arg Cys Leu Ala Glu Asn Ser Leu Gly Ser 305
310 315 320 Ala Arg His Ala
Tyr Tyr Val Thr Val Glu Ala Ala Pro Tyr Trp Leu 325
330 335 His Lys Pro Gln Ser His Leu Tyr Gly
Pro Gly Glu Thr Ala Arg Leu 340 345
350 Asp Cys Gln Val Gln Gly Arg Pro Gln Pro Glu Val Thr Trp
Arg Ile 355 360 365
Asn Gly Ile Pro Val Glu Glu Leu Ala Lys Asp Gln Lys Tyr Arg Ile 370
375 380 Gln Arg Gly Ala Leu
Ile Leu Ser Asn Val Gln Pro Ser Asp Thr Met 385 390
395 400 Val Thr Gln Cys Glu Ala Arg Asn Arg His
Gly Leu Leu Leu Ala Asn 405 410
415 Ala Tyr Ile Tyr Val Val Gln Leu Pro Ala Lys Ile Leu Thr Ala
Asp 420 425 430 Asn
Gln Thr Tyr Met Ala Val Gln Gly Ser Thr Ala Tyr Leu Leu Cys 435
440 445 Lys Ala Phe Gly Ala Pro
Val Pro Ser Val Gln Trp Leu Asp Glu Asp 450 455
460 Gly Thr Thr Val Leu Gln Asp Glu Arg Phe Phe
Pro Tyr Ala Asn Gly 465 470 475
480 Thr Leu Gly Ile Arg Asp Leu Gln Ala Asn Asp Thr Gly Arg Tyr Phe
485 490 495 Cys Leu
Ala Ala Asn Asp Gln Asn Asn Val Thr Ile Met Ala Asn Leu 500
505 510 Lys Val Lys Asp Ala Thr Gln
Ile Thr Gln Gly Pro Arg Ser Thr Ile 515 520
525 Glu Lys Lys Gly Ser Arg Val Thr Phe Thr Cys Gln
Ala Ser Phe Asp 530 535 540
Pro Ser Leu Gln Pro Ser Ile Thr Trp Arg Gly Asp Gly Arg Asp Leu 545
550 555 560 Gln Glu Leu
Gly Asp Ser Asp Lys Tyr Phe Ile Glu Asp Gly Arg Leu 565
570 575 Val Ile His Ser Leu Asp Tyr Ser
Asp Gln Gly Asn Tyr Ser Cys Val 580 585
590 Ala Ser Thr Glu Leu Asp Val Val Glu Ser Arg Ala Gln
Leu Leu Val 595 600 605
Val Gly Ser Pro Gly Pro Val Pro Arg Leu Val Leu Ser Asp Leu His 610
615 620 Leu Leu Thr Gln
Ser Gln Val Arg Val Ser Trp Ser Pro Ala Glu Asp 625 630
635 640 His Asn Ala Pro Ile Glu Lys Tyr Asp
Ile Glu Phe Glu Asp Lys Glu 645 650
655 Met Ala Pro Glu Lys Trp Tyr Ser Leu Gly Lys Val Pro Gly
Asn Gln 660 665 670
Thr Ser Thr Thr Leu Lys Leu Ser Pro Tyr Val His Tyr Thr Phe Arg
675 680 685 Val Thr Ala Ile
Asn Lys Tyr Gly Pro Gly Glu Pro Ser Pro Val Ser 690
695 700 Glu Thr Val Val Thr Pro Glu Ala
Ala Pro Glu Lys Asn Pro Val Asp 705 710
715 720 Val Lys Gly Glu Gly Asn Glu Thr Thr Asn Met Val
Ile Thr Trp Lys 725 730
735 Pro Leu Arg Trp Met Asp Trp Asn Ala Pro Gln Val Gln Tyr Arg Val
740 745 750 Gln Trp Arg
Pro Gln Gly Thr Arg Gly Pro Trp Gln Glu Gln Ile Val 755
760 765 Ser Asp Pro Phe Leu Val Val Ser
Asn Thr Ser Thr Phe Val Pro Tyr 770 775
780 Glu Ile Lys Val Gln Ala Val Asn Ser Gln Gly Lys Gly
Pro Glu Pro 785 790 795
800 Gln Val Thr Ile Gly Tyr Ser Gly Glu Asp Tyr Pro Gln Ala Ile Pro
805 810 815 Glu Leu Glu Gly
Ile Glu Ile Leu Asn Ser Ser Ala Val Leu Val Lys 820
825 830 Trp Arg Pro Val Asp Leu Ala Gln Val
Lys Gly His Leu Arg Gly Tyr 835 840
845 Asn Val Thr Tyr Trp Arg Glu Gly Ser Gln Arg Lys His Ser
Lys Arg 850 855 860
His Ile His Lys Asp His Val Val Val Pro Ala Asn Thr Thr Ser Val 865
870 875 880 Ile Leu Ser Gly Leu
Arg Pro Tyr Ser Ser Tyr His Leu Glu Val Gln 885
890 895 Ala Phe Asn Gly Arg Gly Ser Gly Pro Ala
Ser Glu Phe Thr Phe Ser 900 905
910 Thr Pro Glu Gly Val Pro Gly His Pro Glu Ala Leu His Leu Glu
Cys 915 920 925 Gln
Ser Asn Thr Ser Leu Leu Leu Arg Trp Gln Pro Pro Leu Ser His 930
935 940 Asn Gly Val Leu Thr Gly
Tyr Val Leu Ser Tyr His Pro Leu Asp Glu 945 950
955 960 Gly Gly Lys Gly Gln Leu Ser Phe Asn Leu Arg
Asp Pro Glu Leu Arg 965 970
975 Thr His Asn Leu Thr Asp Leu Ser Pro His Leu Arg Tyr Arg Phe Gln
980 985 990 Leu Gln
Ala Thr Thr Lys Glu Gly Pro Gly Glu Ala Ile Val Arg Glu 995
1000 1005 Gly Gly Thr Met Ala
Leu Ser Gly Ile Ser Asp Phe Gly Asn Ile 1010 1015
1020 Ser Ala Thr Ala Gly Glu Asn Tyr Ser Val
Val Ser Trp Val Pro 1025 1030 1035
Lys Glu Gly Gln Cys Asn Phe Arg Phe His Ile Leu Phe Lys Ala
1040 1045 1050 Leu Gly
Glu Glu Lys Gly Gly Ala Ser Leu Ser Pro Gln Tyr Val 1055
1060 1065 Ser Tyr Asn Gln Ser Ser Tyr
Thr Gln Trp Asp Leu Gln Pro Asp 1070 1075
1080 Thr Asp Tyr Glu Ile His Leu Phe Lys Glu Arg Met
Phe Arg His 1085 1090 1095
Gln Met Ala Val Lys Thr Asn Gly Thr Gly Arg Val Arg Leu Pro 1100
1105 1110 Pro Ala Gly Phe Ala
Thr Glu Gly Trp Phe Ile Gly Phe Val Ser 1115 1120
1125 Ala Ile Ile Leu Leu Leu Leu Val Leu Leu
Ile Leu Cys Phe Ile 1130 1135 1140
Lys Arg Ser Lys Gly Gly Lys Tyr Ser Val Lys Asp Lys Glu Asp
1145 1150 1155 Thr Gln
Val Asp Ser Glu Ala Arg Pro Met Lys Asp Glu Thr Phe 1160
1165 1170 Gly Glu Tyr Arg Ser Leu Glu
Ser Asp Asn Glu Glu Lys Ala Phe 1175 1180
1185 Gly Ser Ser Gln Pro Ser Leu Asn Gly Asp Ile Lys
Pro Leu Gly 1190 1195 1200
Ser Asp Asp Ser Leu Ala Asp Tyr Gly Gly Ser Val Asp Val Gln 1205
1210 1215 Phe Asn Glu Asp Gly
Ser Phe Ile Gly Gln Tyr Ser Gly Lys Lys 1220 1225
1230 Glu Lys Glu Ala Ala Gly Gly Asn Asp Ser
Ser Gly Ala Thr Ser 1235 1240 1245
Pro Ile Asn Pro Ala Val Ala Leu Glu 1250
1255 6640PRTUnknownHomo sapiens GGF2 protein 6Met Ser Glu Arg Lys
Glu Gly Arg Gly Lys Gly Lys Gly Lys Lys Lys 1 5
10 15 Glu Arg Gly Ser Gly Lys Lys Pro Glu Ser
Ala Ala Gly Ser Gln Ser 20 25
30 Pro Ala Leu Pro Pro Arg Leu Lys Glu Met Lys Ser Gln Glu Ser
Ala 35 40 45 Ala
Gly Ser Lys Leu Val Leu Arg Cys Glu Thr Ser Ser Glu Tyr Ser 50
55 60 Ser Leu Arg Phe Lys Trp
Phe Lys Asn Gly Asn Glu Leu Asn Arg Lys 65 70
75 80 Asn Lys Pro Gln Asn Ile Lys Ile Gln Lys Lys
Pro Gly Lys Ser Glu 85 90
95 Leu Arg Ile Asn Lys Ala Ser Leu Ala Asp Ser Gly Glu Tyr Met Cys
100 105 110 Lys Val
Ile Ser Lys Leu Gly Asn Asp Ser Ala Ser Ala Asn Ile Thr 115
120 125 Ile Val Glu Ser Asn Glu Ile
Ile Thr Gly Met Pro Ala Ser Thr Glu 130 135
140 Gly Ala Tyr Val Ser Ser Glu Ser Pro Ile Arg Ile
Ser Val Ser Thr 145 150 155
160 Glu Gly Ala Asn Thr Ser Ser Ser Thr Ser Thr Ser Thr Thr Gly Thr
165 170 175 Ser His Leu
Val Lys Cys Ala Glu Lys Glu Lys Thr Phe Cys Val Asn 180
185 190 Gly Gly Glu Cys Phe Met Val Lys
Asp Leu Ser Asn Pro Ser Arg Tyr 195 200
205 Leu Cys Lys Cys Gln Pro Gly Phe Thr Gly Ala Arg Cys
Thr Glu Asn 210 215 220
Val Pro Met Lys Val Gln Asn Gln Glu Lys Ala Glu Glu Leu Tyr Gln 225
230 235 240 Lys Arg Val Leu
Thr Ile Thr Gly Ile Cys Ile Ala Leu Leu Val Val 245
250 255 Gly Ile Met Cys Val Val Ala Tyr Cys
Lys Thr Lys Lys Gln Arg Lys 260 265
270 Lys Leu His Asp Arg Leu Arg Gln Ser Leu Arg Ser Glu Arg
Asn Asn 275 280 285
Met Met Asn Ile Ala Asn Gly Pro His His Pro Asn Pro Pro Pro Glu 290
295 300 Asn Val Gln Leu Val
Asn Gln Tyr Val Ser Lys Asn Val Ile Ser Ser 305 310
315 320 Glu His Ile Val Glu Arg Glu Ala Glu Thr
Ser Phe Ser Thr Ser His 325 330
335 Tyr Thr Ser Thr Ala His His Ser Thr Thr Val Thr Gln Thr Pro
Ser 340 345 350 His
Ser Trp Ser Asn Gly His Thr Glu Ser Ile Leu Ser Glu Ser His 355
360 365 Ser Val Ile Val Met Ser
Ser Val Glu Asn Ser Arg His Ser Ser Pro 370 375
380 Thr Gly Gly Pro Arg Gly Arg Leu Asn Gly Thr
Gly Gly Pro Arg Glu 385 390 395
400 Cys Asn Ser Phe Leu Arg His Ala Arg Glu Thr Pro Asp Ser Tyr Arg
405 410 415 Asp Ser
Pro His Ser Glu Arg Tyr Val Ser Ala Met Thr Thr Pro Ala 420
425 430 Arg Met Ser Pro Val Asp Phe
His Thr Pro Ser Ser Pro Lys Ser Pro 435 440
445 Pro Ser Glu Met Ser Pro Pro Val Ser Ser Met Thr
Val Ser Met Pro 450 455 460
Ser Met Ala Val Ser Pro Phe Met Glu Glu Glu Arg Pro Leu Leu Leu 465
470 475 480 Val Thr Pro
Pro Arg Leu Arg Glu Lys Lys Phe Asp His His Pro Gln 485
490 495 Gln Phe Ser Ser Phe His His Asn
Pro Ala His Asp Ser Asn Ser Leu 500 505
510 Pro Ala Ser Pro Leu Arg Ile Val Glu Asp Glu Glu Tyr
Glu Thr Thr 515 520 525
Gln Glu Tyr Glu Pro Ala Gln Glu Pro Val Lys Lys Leu Ala Asn Ser 530
535 540 Arg Arg Ala Lys
Arg Thr Lys Pro Asn Gly His Ile Ala Asn Arg Leu 545 550
555 560 Glu Val Asp Ser Asn Thr Ser Ser Gln
Ser Ser Asn Ser Glu Ser Glu 565 570
575 Thr Glu Asp Glu Arg Val Gly Glu Asp Thr Pro Phe Leu Gly
Ile Gln 580 585 590
Asn Pro Leu Ala Ala Ser Leu Glu Ala Thr Pro Ala Phe Arg Leu Ala
595 600 605 Asp Ser Arg Thr
Asn Pro Ala Gly Arg Phe Ser Thr Gln Glu Glu Ile 610
615 620 Gln Ala Arg Leu Ser Ser Val Ile
Ala Asn Gln Asp Pro Ile Ala Val 625 630
635 640 7199PRTUnknownHomo sapiens Brain derived
neurotrophic factor protein 7Met Thr Ile Leu Phe Leu Thr Met Val Ile
Ser Tyr Phe Gly Cys Met 1 5 10
15 Lys Ala Ala Pro Met Lys Glu Ala Asn Ile Arg Gly Gln Gly Gly
Leu 20 25 30 Ala
Tyr Pro Gly Val Arg Thr His Gly Thr Leu Glu Ser Val Asn Gly 35
40 45 Pro Lys Ala Gly Ser Arg
Gly Leu Thr Ser Leu Ala Asp Thr Phe Glu 50 55
60 His Val Ile Glu Glu Leu Leu Asp Glu Asp Gln
Lys Val Arg Pro Asn 65 70 75
80 Glu Glu Asn Asn Lys Asp Ala Asp Leu Tyr Thr Ser Arg Val Met Leu
85 90 95 Ser Ser
Gln Val Pro Leu Glu Pro Pro Leu Leu Phe Leu Leu Glu Glu 100
105 110 Tyr Lys Asn Tyr Leu Asp Ala
Ala Asn Met Ser Met Arg Val Arg Arg 115 120
125 His Ser Asp Pro Ala Arg Arg Gly Glu Leu Ser Val
Cys Asp Ser Ile 130 135 140
Ser Glu Trp Val Thr Ala Ala Asp Lys Arg His Trp Asn Ser Gln Cys 145
150 155 160 Arg Thr Thr
Gln Ser Tyr Val Arg Ala Leu Thr Met Asp Ser Lys Lys 165
170 175 Arg Ile Gly Trp Arg Phe Ile Arg
Ile Asp Thr Ser Cys Val Cys Thr 180 185
190 Leu Thr Ile Lys Arg Gly Arg 195
8282PRTUnknownHomo sapiens Insulin-like growth factor protein 8Met
Glu Arg Pro Ser Leu Arg Ala Leu Leu Leu Gly Ala Ala Gly Leu 1
5 10 15 Leu Leu Leu Leu Leu Pro
Leu Ser Ser Ser Ser Ser Ser Asp Thr Cys 20
25 30 Gly Pro Cys Glu Pro Ala Ser Cys Pro Pro
Leu Pro Pro Leu Gly Cys 35 40
45 Leu Leu Gly Glu Thr Arg Asp Ala Cys Gly Cys Cys Pro Met
Cys Ala 50 55 60
Arg Gly Glu Gly Glu Pro Cys Gly Gly Gly Gly Ala Gly Arg Gly Tyr 65
70 75 80 Cys Ala Pro Gly Met
Glu Cys Val Lys Ser Arg Lys Arg Arg Lys Gly 85
90 95 Lys Ala Gly Ala Ala Ala Gly Gly Pro Gly
Val Ser Gly Val Cys Val 100 105
110 Cys Lys Ser Arg Tyr Pro Val Cys Gly Ser Asp Gly Thr Thr Tyr
Pro 115 120 125 Ser
Gly Cys Gln Leu Arg Ala Ala Ser Gln Arg Ala Glu Ser Arg Gly 130
135 140 Glu Lys Ala Ile Thr Gln
Val Ser Lys Gly Thr Cys Glu Gln Gly Pro 145 150
155 160 Ser Ile Val Thr Pro Pro Lys Asp Ile Trp Asn
Val Thr Gly Ala Gln 165 170
175 Val Tyr Leu Ser Cys Glu Val Ile Gly Ile Pro Thr Pro Val Leu Ile
180 185 190 Trp Asn
Lys Val Lys Arg Gly His Tyr Gly Val Gln Arg Thr Glu Leu 195
200 205 Leu Pro Gly Asp Arg Asp Asn
Leu Ala Ile Gln Thr Arg Gly Gly Pro 210 215
220 Glu Lys His Glu Val Thr Gly Trp Val Leu Val Ser
Pro Leu Ser Lys 225 230 235
240 Glu Asp Ala Gly Glu Tyr Glu Cys His Ala Ser Asn Ser Gln Gly Gln
245 250 255 Ala Ser Ala
Ser Ala Lys Ile Thr Val Val Asp Ala Leu His Glu Ile 260
265 270 Pro Val Lys Lys Gly Glu Gly Ala
Glu Leu 275 280 9155PRTUnknownHomo
sapiens Fibroblast growth factor protein 9Met Ala Glu Gly Glu Ile Thr Thr
Phe Thr Ala Leu Thr Glu Lys Phe 1 5 10
15 Asn Leu Pro Pro Gly Asn Tyr Lys Lys Pro Lys Leu Leu
Tyr Cys Ser 20 25 30
Asn Gly Gly His Phe Leu Arg Ile Leu Pro Asp Gly Thr Val Asp Gly
35 40 45 Thr Arg Asp Arg
Ser Asp Gln His Ile Gln Leu Gln Leu Ser Ala Glu 50
55 60 Ser Val Gly Glu Val Tyr Ile Lys
Ser Thr Glu Thr Gly Gln Tyr Leu 65 70
75 80 Ala Met Asp Thr Asp Gly Leu Leu Tyr Gly Ser Gln
Thr Pro Asn Glu 85 90
95 Glu Cys Leu Phe Leu Glu Arg Leu Glu Glu Asn His Tyr Asn Thr Tyr
100 105 110 Ile Ser Lys
Lys His Ala Glu Lys Asn Trp Phe Val Gly Leu Lys Lys 115
120 125 Asn Gly Ser Cys Lys Arg Gly Pro
Arg Thr His Tyr Gly Gln Lys Ala 130 135
140 Ile Leu Phe Leu Pro Leu Pro Val Ser Ser Asp 145
150 155 10200PRTUnknownHomo sapiens Ciliary
neurotrophic factor protein 10Met Ala Phe Thr Glu His Ser Pro Leu Thr Pro
His Arg Arg Asp Leu 1 5 10
15 Cys Ser Arg Ser Ile Trp Leu Ala Arg Lys Ile Arg Ser Asp Leu Thr
20 25 30 Ala Leu
Thr Glu Ser Tyr Val Lys His Gln Gly Leu Asn Lys Asn Ile 35
40 45 Asn Leu Asp Ser Ala Asp Gly
Met Pro Val Ala Ser Thr Asp Gln Trp 50 55
60 Ser Glu Leu Thr Glu Ala Glu Arg Leu Gln Glu Asn
Leu Gln Ala Tyr 65 70 75
80 Arg Thr Phe His Val Leu Leu Ala Arg Leu Leu Glu Asp Gln Gln Val
85 90 95 His Phe Thr
Pro Thr Glu Gly Asp Phe His Gln Ala Ile His Thr Leu 100
105 110 Leu Leu Gln Val Ala Ala Phe Ala
Tyr Gln Ile Glu Glu Leu Met Ile 115 120
125 Leu Leu Glu Tyr Lys Ile Pro Arg Asn Glu Ala Asp Gly
Met Pro Ile 130 135 140
Asn Val Gly Asp Gly Gly Leu Phe Glu Lys Lys Leu Trp Gly Leu Lys 145
150 155 160 Val Leu Gln Glu
Leu Ser Gln Trp Thr Val Arg Ser Ile His Asp Leu 165
170 175 Arg Phe Ile Ser Ser His Gln Thr Gly
Ile Pro Ala Arg Gly Ser His 180 185
190 Tyr Ile Ala Asn Asn Lys Lys Met 195
200 11211PRTUnknownHomo sapiens Glial derived neurotrophic factor
protein 11Met Lys Leu Trp Asp Val Val Ala Val Cys Leu Val Leu Leu His
Thr 1 5 10 15 Ala
Ser Ala Phe Pro Leu Pro Ala Gly Lys Arg Pro Pro Glu Ala Pro
20 25 30 Ala Glu Asp Arg Ser
Leu Gly Arg Arg Arg Ala Pro Phe Ala Leu Ser 35
40 45 Ser Asp Ser Asn Met Pro Glu Asp Tyr
Pro Asp Gln Phe Asp Asp Val 50 55
60 Met Asp Phe Ile Gln Ala Thr Ile Lys Arg Leu Lys Arg
Ser Pro Asp 65 70 75
80 Lys Gln Met Ala Val Leu Pro Arg Arg Glu Arg Asn Arg Gln Ala Ala
85 90 95 Ala Ala Asn Pro
Glu Asn Ser Arg Gly Lys Gly Arg Arg Gly Gln Arg 100
105 110 Gly Lys Asn Arg Gly Cys Val Leu Thr
Ala Ile His Leu Asn Val Thr 115 120
125 Asp Leu Gly Leu Gly Tyr Glu Thr Lys Glu Glu Leu Ile Phe
Arg Tyr 130 135 140
Cys Ser Gly Ser Cys Asp Ala Ala Glu Thr Thr Tyr Asp Lys Ile Leu 145
150 155 160 Lys Asn Leu Ser Arg
Asn Arg Arg Leu Val Ser Asp Lys Val Gly Gln 165
170 175 Ala Cys Cys Arg Pro Ile Ala Phe Asp Asp
Asp Leu Ser Phe Leu Asp 180 185
190 Asp Asn Leu Val Tyr His Ile Leu Arg Lys His Ser Ala Lys Arg
Cys 195 200 205 Gly
Cys Ile 210 12414PRTUnknownHomo sapiens Transforming growth
factor protein 12Met His Tyr Cys Val Leu Ser Ala Phe Leu Ile Leu His Leu
Val Thr 1 5 10 15
Val Ala Leu Ser Leu Ser Thr Cys Ser Thr Leu Asp Met Asp Gln Phe
20 25 30 Met Arg Lys Arg Ile
Glu Ala Ile Arg Gly Gln Ile Leu Ser Lys Leu 35
40 45 Lys Leu Thr Ser Pro Pro Glu Asp Tyr
Pro Glu Pro Glu Glu Val Pro 50 55
60 Pro Glu Val Ile Ser Ile Tyr Asn Ser Thr Arg Asp Leu
Leu Gln Glu 65 70 75
80 Lys Ala Ser Arg Arg Ala Ala Ala Cys Glu Arg Glu Arg Ser Asp Glu
85 90 95 Glu Tyr Tyr Ala
Lys Glu Val Tyr Lys Ile Asp Met Pro Pro Phe Phe 100
105 110 Pro Ser Glu Asn Ala Ile Pro Pro Thr
Phe Tyr Arg Pro Tyr Phe Arg 115 120
125 Ile Val Arg Phe Asp Val Ser Ala Met Glu Lys Asn Ala Ser
Asn Leu 130 135 140
Val Lys Ala Glu Phe Arg Val Phe Arg Leu Gln Asn Pro Lys Ala Arg 145
150 155 160 Val Pro Glu Gln Arg
Ile Glu Leu Tyr Gln Ile Leu Lys Ser Lys Asp 165
170 175 Leu Thr Ser Pro Thr Gln Arg Tyr Ile Asp
Ser Lys Val Val Lys Thr 180 185
190 Arg Ala Glu Gly Glu Trp Leu Ser Phe Asp Val Thr Asp Ala Val
His 195 200 205 Glu
Trp Leu His His Lys Asp Arg Asn Leu Gly Phe Lys Ile Ser Leu 210
215 220 His Cys Pro Cys Cys Thr
Phe Val Pro Ser Asn Asn Tyr Ile Ile Pro 225 230
235 240 Asn Lys Ser Glu Glu Leu Glu Ala Arg Phe Ala
Gly Ile Asp Gly Thr 245 250
255 Ser Thr Tyr Thr Ser Gly Asp Gln Lys Thr Ile Lys Ser Thr Arg Lys
260 265 270 Lys Asn
Ser Gly Lys Thr Pro His Leu Leu Leu Met Leu Leu Pro Ser 275
280 285 Tyr Arg Leu Glu Ser Gln Gln
Thr Asn Arg Arg Lys Lys Arg Ala Leu 290 295
300 Asp Ala Ala Tyr Cys Phe Arg Asn Val Gln Asp Asn
Cys Cys Leu Arg 305 310 315
320 Pro Leu Tyr Ile Asp Phe Lys Arg Asp Leu Gly Trp Lys Trp Ile His
325 330 335 Glu Pro Lys
Gly Tyr Asn Ala Asn Phe Cys Ala Gly Ala Cys Pro Tyr 340
345 350 Leu Trp Ser Ser Asp Thr Gln His
Ser Arg Val Leu Ser Leu Tyr Asn 355 360
365 Thr Ile Asn Pro Glu Ala Ser Ala Ser Pro Cys Cys Val
Ser Gln Asp 370 375 380
Leu Glu Pro Leu Thr Ile Leu Tyr Tyr Ile Gly Lys Thr Pro Lys Ile 385
390 395 400 Glu Gln Leu Ser
Asn Met Ile Val Lys Ser Cys Lys Cys Ser 405
410 131021PRTUnknownHomo sapiens Chondroitinase ABC I
protein 13Met Pro Ile Phe Arg Phe Thr Ala Leu Ala Met Thr Leu Gly Leu Leu
1 5 10 15 Ser Ala
Pro Tyr Asn Ala Met Ala Ala Thr Ser Asn Pro Ala Phe Asp 20
25 30 Pro Lys Asn Leu Met Gln Ser
Glu Ile Tyr His Phe Ala Gln Asn Asn 35 40
45 Pro Leu Ala Asp Phe Ser Ser Asp Lys Asn Ser Ile
Leu Thr Leu Ser 50 55 60
Asp Lys Arg Ser Ile Met Gly Asn Gln Ser Leu Leu Trp Lys Trp Lys 65
70 75 80 Gly Gly Ser
Ser Phe Thr Leu His Lys Lys Leu Ile Val Pro Thr Asp 85
90 95 Lys Glu Ala Ser Lys Ala Trp Gly
Arg Ser Ser Thr Pro Val Phe Ser 100 105
110 Phe Trp Leu Tyr Asn Glu Lys Pro Ile Asp Gly Tyr Leu
Thr Ile Asp 115 120 125
Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala Gln Ala Gly Phe Lys 130
135 140 Val Lys Leu Asp
Phe Thr Gly Trp Arg Ala Val Gly Val Ser Leu Asn 145 150
155 160 Asn Asp Leu Glu Asn Arg Glu Met Thr
Leu Asn Ala Thr Asn Thr Ser 165 170
175 Ser Asp Gly Thr Gln Asp Ser Ile Gly Arg Ser Leu Gly Ala
Lys Val 180 185 190
Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn Val Ser Gln Gly Glu Ile
195 200 205 Tyr Ile Asp Arg
Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp 210
215 220 Ser Asp Tyr Gln Val Lys Thr Arg
Leu Ser Glu Pro Glu Ile Gln Phe 225 230
235 240 His Asn Val Lys Pro Gln Leu Pro Val Thr Pro Glu
Asn Leu Ala Ala 245 250
255 Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu Phe Val Gly Gly Glu
260 265 270 Lys Glu Thr
Asn Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser 275
280 285 Asp Phe Asp Ala Leu Asn Ile His
Thr Leu Ala Asn Gly Gly Thr Gln 290 295
300 Gly Arg His Leu Ile Thr Asp Lys Gln Ile Ile Ile Tyr
Gln Pro Glu 305 310 315
320 Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp Asn Tyr Val Ile Leu
325 330 335 Gly Asn Tyr Thr
Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val Leu 340
345 350 Glu Lys Asp Pro Thr Gln Lys Ala Gln
Leu Lys Gln Met Tyr Leu Leu 355 360
365 Met Thr Lys His Leu Leu Asp Gln Gly Phe Val Lys Gly Ser
Ala Leu 370 375 380
Val Thr Thr His His Trp Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser 385
390 395 400 Thr Leu Leu Met Ser
Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln 405
410 415 Val Tyr Asp Ser Leu Leu Trp Tyr Ser Arg
Glu Phe Lys Ser Ser Phe 420 425
430 Asp Met Lys Val Ser Ala Asp Ser Ser Asp Leu Asp Tyr Phe Asn
Thr 435 440 445 Leu
Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln 450
455 460 Lys Arg Ile Asn Leu Val
Asn Thr Phe Ser His Tyr Ile Thr Gly Ala 465 470
475 480 Leu Thr Gln Val Pro Pro Gly Gly Lys Asp Gly
Leu Arg Pro Asp Gly 485 490
495 Thr Ala Trp Arg His Glu Gly Asn Tyr Pro Gly Tyr Ser Phe Pro Ala
500 505 510 Phe Lys
Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe 515
520 525 Ser Val Gly Glu Ser Gly Trp
Asn Asn Leu Lys Lys Ala Met Val Ser 530 535
540 Ala Trp Ile Tyr Ser Asn Pro Glu Val Gly Leu Pro
Leu Ala Gly Arg 545 550 555
560 His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val Ala Gln Gly Tyr Tyr
565 570 575 Trp Leu Ala
Met Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser 580
585 590 Ile Tyr Leu Ala Ile Ser Asp Lys
Thr Gln Asn Glu Ser Thr Ala Ile 595 600
605 Phe Gly Glu Thr Ile Thr Pro Ala Ser Leu Pro Gln Gly
Phe Tyr Ala 610 615 620
Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met Val 625
630 635 640 Thr Leu Lys Ala
Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn 645
650 655 Lys Asp Asn Arg Tyr Gly Arg Tyr Gln
Ser His Gly Val Ala Gln Ile 660 665
670 Val Ser Asn Gly Ser Gln Leu Ser Gln Gly Tyr Gln Gln Glu
Gly Trp 675 680 685
Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys 690
695 700 Asp Leu Asp Ser Pro
Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg 705 710
715 720 Gly Phe Ser Gly Thr Ser Ser Leu Glu Gly
Gln Tyr Gly Met Met Ala 725 730
735 Phe Asn Leu Ile Tyr Pro Ala Asn Leu Glu Arg Phe Asp Pro Asn
Phe 740 745 750 Thr
Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile 755
760 765 Gly Ser Asn Ile Asn Ser
Ser Asp Lys Asn Lys Asn Val Glu Thr Thr 770 775
780 Leu Phe Gln His Ala Ile Thr Pro Thr Leu Asn
Thr Leu Trp Ile Asn 785 790 795
800 Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln Thr Thr Leu Gln Gln Gly
805 810 815 Asp Trp
Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala 820
825 830 Glu Lys Val Asn Val Ser Arg
Gln His Gln Val Ser Ala Glu Asn Lys 835 840
845 Asn Arg Gln Pro Thr Glu Gly Asn Phe Ser Ser Ala
Trp Ile Asp His 850 855 860
Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr Met Val Phe Leu Asp 865
870 875 880 Ala Thr Pro
Glu Lys Met Gly Glu Met Ala Gln Lys Phe Arg Glu Asn 885
890 895 Asn Gly Leu Tyr Gln Val Leu Arg
Lys Asp Lys Asp Val His Ile Ile 900 905
910 Leu Asp Lys Leu Ser Asn Val Thr Gly Tyr Ala Phe Tyr
Gln Pro Ala 915 920 925
Ser Ile Glu Asp Lys Trp Ile Lys Lys Val Asn Lys Pro Ala Ile Val 930
935 940 Met Thr His Arg
Gln Lys Asp Thr Leu Ile Val Ser Ala Val Thr Pro 945 950
955 960 Asp Leu Asn Met Thr Arg Gln Lys Ala
Ala Thr Pro Val Thr Ile Asn 965 970
975 Val Thr Ile Asn Gly Lys Trp Gln Ser Ala Asp Lys Asn Ser
Glu Val 980 985 990
Lys Tyr Gln Val Ser Gly Asp Asn Thr Glu Leu Thr Phe Thr Ser Tyr
995 1000 1005 Phe Gly Ile
Pro Gln Glu Ile Lys Leu Ser Pro Leu Pro 1010 1015
1020 14232PRTUnknownHomo sapiens Vascular endothelial
growth factor protein 14Met Asn Phe Leu Leu Ser Trp Val His Trp Ser
Leu Ala Leu Leu Leu 1 5 10
15 Tyr Leu His His Ala Lys Trp Ser Gln Ala Ala Pro Met Ala Glu Gly
20 25 30 Gly Gly
Gln Asn His His Glu Val Val Lys Phe Met Asp Val Tyr Gln 35
40 45 Arg Ser Tyr Cys His Pro Ile
Glu Thr Leu Val Asp Ile Phe Gln Glu 50 55
60 Tyr Pro Asp Glu Ile Glu Tyr Ile Phe Lys Pro Ser
Cys Val Pro Leu 65 70 75
80 Met Arg Cys Gly Gly Cys Cys Asn Asp Glu Gly Leu Glu Cys Val Pro
85 90 95 Thr Glu Glu
Ser Asn Ile Thr Met Gln Ile Met Arg Ile Lys Pro His 100
105 110 Gln Gly Gln His Ile Gly Glu Met
Ser Phe Leu Gln His Asn Lys Cys 115 120
125 Glu Cys Arg Pro Lys Lys Asp Arg Ala Arg Gln Glu Lys
Lys Ser Val 130 135 140
Arg Gly Lys Gly Lys Gly Gln Lys Arg Lys Arg Lys Lys Ser Arg Tyr 145
150 155 160 Lys Ser Trp Ser
Val Tyr Val Gly Ala Arg Cys Cys Leu Met Pro Trp 165
170 175 Ser Leu Pro Gly Pro His Pro Cys Gly
Pro Cys Ser Glu Arg Arg Lys 180 185
190 His Leu Phe Val Gln Asp Pro Gln Thr Cys Lys Cys Ser Cys
Lys Asn 195 200 205
Thr Asp Ser Arg Cys Lys Ala Arg Gln Leu Glu Leu Asn Glu Arg Thr 210
215 220 Cys Arg Cys Asp Lys
Pro Arg Arg 225 230 15240PRTUnknownHomo sapiens
Nerve growth factor protein 15Met Ser Met Leu Phe Tyr Thr Leu Ile Thr Ala
Phe Leu Ile Gly Ile 1 5 10
15 Gln Ala Glu Pro His Ser Glu Ser Asn Val Pro Ala Gly His Thr Ile
20 25 30 Pro Gln
Val His Trp Thr Lys Leu Gln His Ser Leu Asp Thr Ala Leu 35
40 45 Arg Arg Ala Arg Ser Ala Pro
Ala Ala Ala Ile Ala Ala Arg Val Ala 50 55
60 Gly Gln Thr Arg Asn Ile Thr Val Asp Pro Arg Leu
Phe Lys Lys Arg 65 70 75
80 Arg Leu Arg Ser Pro Arg Val Leu Phe Ser Thr Gln Pro Pro Arg Glu
85 90 95 Ala Ala Asp
Thr Gln Asp Leu Asp Phe Glu Val Gly Gly Ala Ala Pro 100
105 110 Phe Asn Arg Thr His Arg Ser Lys
Arg Ser Ser Ser His Pro Ile Phe 115 120
125 His Arg Gly Glu Phe Ser Val Cys Asp Ser Val Ser Val
Trp Val Gly 130 135 140
Asp Lys Thr Thr Ala Thr Asp Ile Lys Gly Lys Glu Val Met Val Leu 145
150 155 160 Gly Glu Val Asn
Ile Asn Asn Ser Val Phe Lys Gln Tyr Phe Phe Glu 165
170 175 Thr Lys Cys Arg Asp Pro Asn Pro Val
Asp Ser Gly Cys Arg Gly Ile 180 185
190 Asp Ser Lys His Trp Asn Ser Tyr Cys Thr Thr Thr His Thr
Phe Val 195 200 205
Lys Ala Leu Thr Met Asp Gly Lys Gln Ala Ala Trp Arg Phe Ile Arg 210
215 220 Ile Asp Thr Ala Cys
Val Cys Val Leu Ser Arg Lys Ala Val Arg Arg 225 230
235 240 16153PRTArtificialSynthetic
polypeptide, GGF2-L(sub)250-C(sub)402 16Cys Ile Ala Leu Leu Val Val Gly
Ile Met Cys Val Val Ala Tyr Cys 1 5 10
15 Lys Thr Lys Lys Gln Arg Lys Lys Leu His Asp Arg Leu
Arg Gln Ser 20 25 30
Leu Arg Ser Glu Arg Asn Asn Met Met Asn Ile Ala Asn Gly Pro His
35 40 45 His Pro Asn Pro
Pro Pro Glu Asn Val Gln Leu Val Asn Gln Tyr Val 50
55 60 Ser Lys Asn Val Ile Ser Ser Glu
His Ile Val Glu Arg Glu Ala Glu 65 70
75 80 Thr Ser Phe Ser Thr Ser His Tyr Thr Ser Thr Ala
His His Ser Thr 85 90
95 Thr Val Thr Gln Thr Pro Ser His Ser Trp Ser Asn Gly His Thr Glu
100 105 110 Ser Ile Leu
Ser Glu Ser His Ser Val Ile Val Met Ser Ser Val Glu 115
120 125 Asn Ser Arg His Ser Ser Pro Thr
Gly Gly Pro Arg Gly Arg Leu Asn 130 135
140 Gly Thr Gly Gly Pro Arg Glu Cys Asn 145
150 17173PRTArtificialSynthetic polypeptide,
GGF2-L(sub)250-E(sub)422 17Cys Ile Ala Leu Leu Val Val Gly Ile Met Cys
Val Val Ala Tyr Cys 1 5 10
15 Lys Thr Lys Lys Gln Arg Lys Lys Leu His Asp Arg Leu Arg Gln Ser
20 25 30 Leu Arg
Ser Glu Arg Asn Asn Met Met Asn Ile Ala Asn Gly Pro His 35
40 45 His Pro Asn Pro Pro Pro Glu
Asn Val Gln Leu Val Asn Gln Tyr Val 50 55
60 Ser Lys Asn Val Ile Ser Ser Glu His Ile Val Glu
Arg Glu Ala Glu 65 70 75
80 Thr Ser Phe Ser Thr Ser His Tyr Thr Ser Thr Ala His His Ser Thr
85 90 95 Thr Val Thr
Gln Thr Pro Ser His Ser Trp Ser Asn Gly His Thr Glu 100
105 110 Ser Ile Leu Ser Glu Ser His Ser
Val Ile Val Met Ser Ser Val Glu 115 120
125 Asn Ser Arg His Ser Ser Pro Thr Gly Gly Pro Arg Gly
Arg Leu Asn 130 135 140
Gly Thr Gly Gly Pro Arg Glu Cys Asn Ser Phe Leu Arg His Ala Arg 145
150 155 160 Glu Thr Pro Asp
Ser Tyr Arg Asp Ser Pro His Ser Glu 165
170 1853PRTArtificialSynthetic polypeptide,
GGF2-T(sub)350-C(sub)402 18Thr Pro Ser His Ser Trp Ser Asn Gly His Thr
Glu Ser Ile Leu Ser 1 5 10
15 Glu Ser His Ser Val Ile Val Met Ser Ser Val Glu Asn Ser Arg His
20 25 30 Ser Ser
Pro Thr Gly Gly Pro Arg Gly Arg Leu Asn Gly Thr Gly Gly 35
40 45 Pro Arg Glu Cys Asn 50
191498PRTArtificialSynthetic polypeptide, N terminal fusion
chimera between ABCI-N (delta)60-C(delta)80 and GGF2-FL 19Met Ser Glu
Arg Lys Glu Gly Arg Gly Lys Gly Lys Gly Lys Lys Lys 1 5
10 15 Glu Arg Gly Ser Gly Lys Lys Pro
Glu Ser Ala Ala Gly Ser Gln Ser 20 25
30 Pro Ala Leu Pro Pro Arg Leu Lys Glu Met Lys Ser Gln
Glu Ser Ala 35 40 45
Ala Gly Ser Lys Leu Val Leu Arg Cys Glu Thr Ser Ser Glu Tyr Ser 50
55 60 Ser Leu Arg Phe
Lys Trp Phe Lys Asn Gly Asn Glu Leu Asn Arg Lys 65 70
75 80 Asn Lys Pro Gln Asn Ile Lys Ile Gln
Lys Lys Pro Gly Lys Ser Glu 85 90
95 Leu Arg Ile Asn Lys Ala Ser Leu Ala Asp Ser Gly Glu Tyr
Met Cys 100 105 110
Lys Val Ile Ser Lys Leu Gly Asn Asp Ser Ala Ser Ala Asn Ile Thr
115 120 125 Ile Val Glu Ser
Asn Glu Ile Ile Thr Gly Met Pro Ala Ser Thr Glu 130
135 140 Gly Ala Tyr Val Ser Ser Glu Ser
Pro Ile Arg Ile Ser Val Ser Thr 145 150
155 160 Glu Gly Ala Asn Thr Ser Ser Ser Thr Ser Thr Ser
Thr Thr Gly Thr 165 170
175 Ser His Leu Val Lys Cys Ala Glu Lys Glu Lys Thr Phe Cys Val Asn
180 185 190 Gly Gly Glu
Cys Phe Met Val Lys Asp Leu Ser Asn Pro Ser Arg Tyr 195
200 205 Leu Cys Lys Cys Gln Pro Gly Phe
Thr Gly Ala Arg Cys Thr Glu Asn 210 215
220 Val Pro Met Lys Val Gln Asn Gln Glu Lys Ala Glu Glu
Leu Tyr Gln 225 230 235
240 Lys Arg Val Leu Thr Ile Thr Gly Ile Cys Ile Ala Leu Leu Val Val
245 250 255 Gly Ile Met Cys
Val Val Ala Tyr Cys Lys Thr Lys Lys Gln Arg Lys 260
265 270 Lys Leu His Asp Arg Leu Arg Gln Ser
Leu Arg Ser Glu Arg Asn Asn 275 280
285 Met Met Asn Ile Ala Asn Gly Pro His His Pro Asn Pro Pro
Pro Glu 290 295 300
Asn Val Gln Leu Val Asn Gln Tyr Val Ser Lys Asn Val Ile Ser Ser 305
310 315 320 Glu His Ile Val Glu
Arg Glu Ala Glu Thr Ser Phe Ser Thr Ser His 325
330 335 Tyr Thr Ser Thr Ala His His Ser Thr Thr
Val Thr Gln Thr Pro Ser 340 345
350 His Ser Trp Ser Asn Gly His Thr Glu Ser Ile Leu Ser Glu Ser
His 355 360 365 Ser
Val Ile Val Met Ser Ser Val Glu Asn Ser Arg His Ser Ser Pro 370
375 380 Thr Gly Gly Pro Arg Gly
Arg Leu Asn Gly Thr Gly Gly Pro Arg Glu 385 390
395 400 Cys Asn Ser Phe Leu Arg His Ala Arg Glu Thr
Pro Asp Ser Tyr Arg 405 410
415 Asp Ser Pro His Ser Glu Arg Tyr Val Ser Ala Met Thr Thr Pro Ala
420 425 430 Arg Met
Ser Pro Val Asp Phe His Thr Pro Ser Ser Pro Lys Ser Pro 435
440 445 Pro Ser Glu Met Ser Pro Pro
Val Ser Ser Met Thr Val Ser Met Pro 450 455
460 Ser Met Ala Val Ser Pro Phe Met Glu Glu Glu Arg
Pro Leu Leu Leu 465 470 475
480 Val Thr Pro Pro Arg Leu Arg Glu Lys Lys Phe Asp His His Pro Gln
485 490 495 Gln Phe Ser
Ser Phe His His Asn Pro Ala His Asp Ser Asn Ser Leu 500
505 510 Pro Ala Ser Pro Leu Arg Ile Val
Glu Asp Glu Glu Tyr Glu Thr Thr 515 520
525 Gln Glu Tyr Glu Pro Ala Gln Glu Pro Val Lys Lys Leu
Ala Asn Ser 530 535 540
Arg Arg Ala Lys Arg Thr Lys Pro Asn Gly His Ile Ala Asn Arg Leu 545
550 555 560 Glu Val Asp Ser
Asn Thr Ser Ser Gln Ser Ser Asn Ser Glu Ser Glu 565
570 575 Thr Glu Asp Glu Arg Val Gly Glu Asp
Thr Pro Phe Leu Gly Ile Gln 580 585
590 Asn Pro Leu Ala Ala Ser Leu Glu Ala Thr Pro Ala Phe Arg
Leu Ala 595 600 605
Asp Ser Arg Thr Asn Pro Ala Gly Arg Phe Ser Thr Gln Glu Glu Ile 610
615 620 Gln Ala Arg Leu Ser
Ser Val Ile Ala Asn Gln Asp Pro Ile Ala Val 625 630
635 640 Phe Thr Leu His Lys Lys Leu Ile Val Pro
Thr Asp Lys Glu Ala Ser 645 650
655 Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu
Tyr 660 665 670 Asn
Glu Lys Pro Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys 675
680 685 Leu Ile Ser Thr Ser Glu
Ala Gln Ala Gly Phe Lys Val Lys Leu Asp 690 695
700 Phe Thr Gly Trp Arg Thr Val Gly Val Ser Leu
Asn Asn Asp Leu Glu 705 710 715
720 Asn Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr
725 730 735 Gln Asp
Ser Ile Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg 740
745 750 Phe Lys Ala Pro Ser Asn Val
Ser Gln Gly Glu Ile Tyr Ile Asp Arg 755 760
765 Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp
Ser Asp Tyr Gln 770 775 780
Val Lys Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys 785
790 795 800 Pro Gln Leu
Pro Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile 805
810 815 Arg Gln Arg Leu Ile Asn Glu Phe
Val Gly Gly Glu Lys Glu Thr Asn 820 825
830 Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser Asp
Phe Asp Ala 835 840 845
Leu Asn Thr His Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu 850
855 860 Ile Thr Asp Lys
Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser 865 870
875 880 Gln Asp Lys Gln Leu Phe Asp Asn Tyr
Val Ile Leu Gly Asn Tyr Thr 885 890
895 Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys
Asp Pro 900 905 910
Thr Gln Lys Ala Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His
915 920 925 Leu Leu Asp Gln
Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His 930
935 940 His Trp Gly Tyr Ser Ser Arg Trp
Trp Tyr Ile Ser Thr Leu Leu Met 945 950
955 960 Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln
Val Tyr Asp Ser 965 970
975 Leu Leu Trp Tyr Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val
980 985 990 Ser Ala Asp
Ser Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln 995
1000 1005 His Leu Ala Leu Leu Leu
Leu Glu Pro Asp Asp Gln Lys Arg Ile 1010 1015
1020 Asn Leu Val Asn Thr Phe Ser His Tyr Ile Thr
Gly Ala Leu Thr 1025 1030 1035
Gln Val Pro Pro Gly Gly Lys Asp Gly Leu Arg Pro Asp Gly Thr
1040 1045 1050 Ala Trp Arg
His Glu Gly Asn Tyr Pro Gly Tyr Ser Phe Pro Ala 1055
1060 1065 Phe Lys Asn Ala Ser Gln Leu Ile
Tyr Leu Leu Arg Asp Thr Pro 1070 1075
1080 Phe Ser Val Gly Glu Ser Gly Trp Asn Asn Leu Lys Lys
Ala Met 1085 1090 1095
Val Ser Ala Trp Ile Tyr Ser Asn Pro Glu Val Gly Leu Pro Leu 1100
1105 1110 Ala Gly Arg His Pro
Phe Asn Ser Pro Ser Leu Lys Ser Val Ala 1115 1120
1125 Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala
Lys Ser Ser Pro Asp 1130 1135 1140
Lys Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser Asp Lys Thr Gln
1145 1150 1155 Asn Glu
Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro Ala Ser 1160
1165 1170 Leu Pro Gln Gly Phe Tyr Ala
Phe Asn Gly Gly Ala Phe Gly Ile 1175 1180
1185 His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala
Tyr Asn Thr 1190 1195 1200
Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg Tyr Gly 1205
1210 1215 Arg Tyr Gln Ser His
Gly Val Ala Gln Ile Val Ser Asn Gly Ser 1220 1225
1230 Gln Leu Ser Gln Gly Tyr Gln Gln Glu Gly
Trp Asp Trp Asn Arg 1235 1240 1245
Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys Asp Leu Asp
1250 1255 1260 Ser Pro
Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg Gly Phe 1265
1270 1275 Ser Gly Thr Ser Ser Leu Glu
Gly Gln Tyr Gly Met Met Ala Phe 1280 1285
1290 Asn Leu Ile Tyr Pro Ala Asn Leu Glu Arg Phe Asp
Pro Asn Phe 1295 1300 1305
Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe 1310
1315 1320 Ile Gly Ser Asn Ile
Asn Ser Ser Asp Lys Asn Lys Asn Val Glu 1325 1330
1335 Thr Thr Leu Phe Gln His Ala Ile Thr Pro
Thr Leu Asn Thr Leu 1340 1345 1350
Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln Thr Thr
1355 1360 1365 Leu Gln
Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr 1370
1375 1380 Leu Ile Thr Gln Ala Glu Lys
Val Asn Val Ser Arg Gln His Gln 1385 1390
1395 Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu
Gly Asn Phe 1400 1405 1410
Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser 1415
1420 1425 Tyr Glu Tyr Met Val
Phe Leu Asp Ala Thr Pro Glu Lys Met Gly 1430 1435
1440 Glu Met Ala Gln Lys Phe Arg Glu Asn Asn
Gly Leu Tyr Gln Val 1445 1450 1455
Leu Arg Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu Ser
1460 1465 1470 Asn Val
Thr Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp 1475
1480 1485 Lys Trp Ile Lys Lys Val Asn
Lys Pro Ala 1490 1495
201011PRTArtificialSynthetic Polypeptide, N Terminal fusion chimera
between ABCI-N(delta) 60-C(delta)80 and GGF2-L(sub)250-C (sub)402
20Cys Ile Ala Leu Leu Val Val Gly Ile Met Cys Val Val Ala Tyr Cys 1
5 10 15 Lys Thr Lys Lys
Gln Arg Lys Lys Leu His Asp Arg Leu Arg Gln Ser 20
25 30 Leu Arg Ser Glu Arg Asn Asn Met Met
Asn Ile Ala Asn Gly Pro His 35 40
45 His Pro Asn Pro Pro Pro Glu Asn Val Gln Leu Val Asn Gln
Tyr Val 50 55 60
Ser Lys Asn Val Ile Ser Ser Glu His Ile Val Glu Arg Glu Ala Glu 65
70 75 80 Thr Ser Phe Ser Thr
Ser His Tyr Thr Ser Thr Ala His His Ser Thr 85
90 95 Thr Val Thr Gln Thr Pro Ser His Ser Trp
Ser Asn Gly His Thr Glu 100 105
110 Ser Ile Leu Ser Glu Ser His Ser Val Ile Val Met Ser Ser Val
Glu 115 120 125 Asn
Ser Arg His Ser Ser Pro Thr Gly Gly Pro Arg Gly Arg Leu Asn 130
135 140 Gly Thr Gly Gly Pro Arg
Glu Cys Asn Phe Thr Leu His Lys Lys Leu 145 150
155 160 Ile Val Pro Thr Asp Lys Glu Ala Ser Lys Ala
Trp Gly Arg Ser Ser 165 170
175 Thr Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys Pro Ile Asp Gly
180 185 190 Tyr Leu
Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala 195
200 205 Gln Ala Gly Phe Lys Val Lys
Leu Asp Phe Thr Gly Trp Arg Thr Val 210 215
220 Gly Val Ser Leu Asn Asn Asp Leu Glu Asn Arg Glu
Met Thr Leu Asn 225 230 235
240 Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile Gly Arg Ser
245 250 255 Leu Gly Ala
Lys Val Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn Val 260
265 270 Ser Gln Gly Glu Ile Tyr Ile Asp
Arg Ile Met Phe Ser Val Asp Asp 275 280
285 Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val Lys Thr Arg
Leu Ser Glu 290 295 300
Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln Leu Pro Val Thr Pro 305
310 315 320 Glu Asn Leu Ala
Ala Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu 325
330 335 Phe Val Gly Gly Glu Lys Glu Thr Asn
Leu Ala Leu Glu Glu Asn Ile 340 345
350 Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr His Thr
Leu Ala 355 360 365
Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp Lys Gln Ile Ile 370
375 380 Ile Tyr Gln Pro Glu
Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp 385 390
395 400 Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr
Leu Met Phe Asn Ile Ser 405 410
415 Arg Ala Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys Ala Gln Leu
Lys 420 425 430 Gln
Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln Gly Phe Val 435
440 445 Lys Gly Ser Ala Leu Val
Thr Thr His His Trp Gly Tyr Ser Ser Arg 450 455
460 Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser Asp
Ala Leu Lys Glu Ala 465 470 475
480 Asn Leu Gln Thr Gln Val Tyr Asp Ser Leu Leu Trp Tyr Ser Arg Glu
485 490 495 Phe Lys
Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser Ser Asp Leu 500
505 510 Asp Tyr Phe Asn Thr Leu Ser
Arg Gln His Leu Ala Leu Leu Leu Leu 515 520
525 Glu Pro Asp Asp Gln Lys Arg Ile Asn Leu Val Asn
Thr Phe Ser His 530 535 540
Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro Gly Gly Lys Asp Gly 545
550 555 560 Leu Arg Pro
Asp Gly Thr Ala Trp Arg His Glu Gly Asn Tyr Pro Gly 565
570 575 Tyr Ser Phe Pro Ala Phe Lys Asn
Ala Ser Gln Leu Ile Tyr Leu Leu 580 585
590 Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly Trp Asn
Asn Leu Lys 595 600 605
Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn Pro Glu Val Gly Leu 610
615 620 Pro Leu Ala Gly
Arg His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val 625 630
635 640 Ala Gln Gly Tyr Tyr Trp Leu Ala Met
Ser Ala Lys Ser Ser Pro Asp 645 650
655 Lys Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser Asp Lys Thr
Gln Asn 660 665 670
Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro Ala Ser Leu Pro
675 680 685 Gln Gly Phe Tyr
Ala Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp 690
695 700 Gln Asp Lys Met Val Thr Leu Lys
Ala Tyr Asn Thr Asn Val Trp Ser 705 710
715 720 Ser Glu Ile Tyr Asn Lys Asp Asn Arg Tyr Gly Arg
Tyr Gln Ser His 725 730
735 Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser Gln Gly Tyr
740 745 750 Gln Gln Glu
Gly Trp Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile 755
760 765 His Leu Pro Leu Lys Asp Leu Asp
Ser Pro Lys Pro His Thr Leu Met 770 775
780 Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr Ser Ser Leu
Glu Gly Gln 785 790 795
800 Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn Leu Glu Arg
805 810 815 Phe Asp Pro Asn
Phe Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn 820
825 830 His Leu Ile Phe Ile Gly Ser Asn Ile
Asn Ser Ser Asp Lys Asn Lys 835 840
845 Asn Val Glu Thr Thr Leu Phe Gln His Ala Ile Thr Pro Thr
Leu Asn 850 855 860
Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln Thr 865
870 875 880 Thr Leu Gln Gln Gly
Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr 885
890 895 Leu Ile Thr Gln Ala Glu Lys Val Asn Val
Ser Arg Gln His Gln Val 900 905
910 Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu Gly Asn Phe Ser
Ser 915 920 925 Ala
Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr 930
935 940 Met Val Phe Leu Asp Ala
Thr Pro Glu Lys Met Gly Glu Met Ala Gln 945 950
955 960 Lys Phe Arg Glu Asn Asn Gly Leu Tyr Gln Val
Leu Arg Lys Asp Lys 965 970
975 Asp Val His Ile Ile Leu Asp Lys Leu Ser Asn Val Thr Gly Tyr Ala
980 985 990 Phe Tyr
Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys Lys Val Asn 995
1000 1005 Lys Pro Ala 1010
211031PRTArtificialSynthetic polypeptide, N terminal fusion
chimera between ABCI-N(delta)60- C(delta)80 and GGF2-L(sub)
250-E(sub)422 21Cys Ile Ala Leu Leu Val Val Gly Ile Met Cys Val Val Ala
Tyr Cys 1 5 10 15
Lys Thr Lys Lys Gln Arg Lys Lys Leu His Asp Arg Leu Arg Gln Ser
20 25 30 Leu Arg Ser Glu Arg
Asn Asn Met Met Asn Ile Ala Asn Gly Pro His 35
40 45 His Pro Asn Pro Pro Pro Glu Asn Val
Gln Leu Val Asn Gln Tyr Val 50 55
60 Ser Lys Asn Val Ile Ser Ser Glu His Ile Val Glu Arg
Glu Ala Glu 65 70 75
80 Thr Ser Phe Ser Thr Ser His Tyr Thr Ser Thr Ala His His Ser Thr
85 90 95 Thr Val Thr Gln
Thr Pro Ser His Ser Trp Ser Asn Gly His Thr Glu 100
105 110 Ser Ile Leu Ser Glu Ser His Ser Val
Ile Val Met Ser Ser Val Glu 115 120
125 Asn Ser Arg His Ser Ser Pro Thr Gly Gly Pro Arg Gly Arg
Leu Asn 130 135 140
Gly Thr Gly Gly Pro Arg Glu Cys Asn Ser Phe Leu Arg His Ala Arg 145
150 155 160 Glu Thr Pro Asp Ser
Tyr Arg Asp Ser Pro His Ser Glu Phe Thr Leu 165
170 175 His Lys Lys Leu Ile Val Pro Thr Asp Lys
Glu Ala Ser Lys Ala Trp 180 185
190 Gly Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu
Lys 195 200 205 Pro
Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser 210
215 220 Thr Ser Glu Ala Gln Ala
Gly Phe Lys Val Lys Leu Asp Phe Thr Gly 225 230
235 240 Trp Arg Thr Val Gly Val Ser Leu Asn Asn Asp
Leu Glu Asn Arg Glu 245 250
255 Met Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp Ser
260 265 270 Ile Gly
Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala 275
280 285 Pro Ser Asn Val Ser Gln Gly
Glu Ile Tyr Ile Asp Arg Ile Met Phe 290 295
300 Ser Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp Tyr
Gln Val Lys Thr 305 310 315
320 Arg Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln Leu
325 330 335 Pro Val Thr
Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg 340
345 350 Leu Ile Asn Glu Phe Val Gly Gly
Glu Lys Glu Thr Asn Leu Ala Leu 355 360
365 Glu Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp Ala
Leu Asn Thr 370 375 380
His Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp 385
390 395 400 Lys Gln Ile Ile
Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys 405
410 415 Gln Leu Phe Asp Asn Tyr Val Ile Leu
Gly Asn Tyr Thr Thr Leu Met 420 425
430 Phe Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro Thr
Gln Lys 435 440 445
Ala Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp 450
455 460 Gln Gly Phe Val Lys
Gly Ser Ala Leu Val Thr Thr His His Trp Gly 465 470
475 480 Tyr Ser Ser Arg Trp Trp Tyr Ile Ser Thr
Leu Leu Met Ser Asp Ala 485 490
495 Leu Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser Leu Leu
Trp 500 505 510 Tyr
Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp 515
520 525 Ser Ser Asp Leu Asp Tyr
Phe Asn Thr Leu Ser Arg Gln His Leu Ala 530 535
540 Leu Leu Leu Leu Glu Pro Asp Asp Gln Lys Arg
Ile Asn Leu Val Asn 545 550 555
560 Thr Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro Gly
565 570 575 Gly Lys
Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly 580
585 590 Asn Tyr Pro Gly Tyr Ser Phe
Pro Ala Phe Lys Asn Ala Ser Gln Leu 595 600
605 Ile Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val Gly
Glu Ser Gly Trp 610 615 620
Asn Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn Pro 625
630 635 640 Glu Val Gly
Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser 645
650 655 Leu Lys Ser Val Ala Gln Gly Tyr
Tyr Trp Leu Ala Met Ser Ala Lys 660 665
670 Ser Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu Ala
Ile Ser Asp 675 680 685
Lys Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro 690
695 700 Ala Ser Leu Pro
Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly 705 710
715 720 Ile His Arg Trp Gln Asp Lys Met Val
Thr Leu Lys Ala Tyr Asn Thr 725 730
735 Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg Tyr
Gly Arg 740 745 750
Tyr Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu
755 760 765 Ser Gln Gly Tyr
Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly 770
775 780 Ala Thr Thr Ile His Leu Pro Leu
Lys Asp Leu Asp Ser Pro Lys Pro 785 790
795 800 His Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser
Gly Thr Ser Ser 805 810
815 Leu Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala
820 825 830 Asn Leu Glu
Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu 835
840 845 Ala Ala Asp Asn His Leu Ile Phe
Ile Gly Ser Asn Ile Asn Ser Ser 850 855
860 Asp Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His
Ala Ile Thr 865 870 875
880 Pro Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met
885 890 895 Pro Tyr Gln Thr
Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn 900
905 910 Gly Asn Gly Tyr Leu Ile Thr Gln Ala
Glu Lys Val Asn Val Ser Arg 915 920
925 Gln His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr
Glu Gly 930 935 940
Asn Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala 945
950 955 960 Ser Tyr Glu Tyr Met
Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly 965
970 975 Glu Met Ala Gln Lys Phe Arg Glu Asn Asn
Gly Leu Tyr Gln Val Leu 980 985
990 Arg Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu Ser
Asn Val 995 1000 1005
Thr Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp 1010
1015 1020 Ile Lys Lys Val Asn
Lys Pro Ala 1025 1030
22911PRTArtificialSynthetic polypeptide, N terminal fusion chimera
between ABCI-N(delta)-C(delta)80 and GGF2-T(sub) 350-C(sub)402 22Thr
Pro Ser His Ser Trp Ser Asn Gly His Thr Glu Ser Ile Leu Ser 1
5 10 15 Glu Ser His Ser Val Ile
Val Met Ser Ser Val Glu Asn Ser Arg His 20
25 30 Ser Ser Pro Thr Gly Gly Pro Arg Gly Arg
Leu Asn Gly Thr Gly Gly 35 40
45 Pro Arg Glu Cys Asn Phe Thr Leu His Lys Lys Leu Ile Val
Pro Thr 50 55 60
Asp Lys Glu Ala Ser Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe 65
70 75 80 Ser Phe Trp Leu Tyr
Asn Glu Lys Pro Ile Asp Gly Tyr Leu Thr Ile 85
90 95 Asp Phe Gly Glu Lys Leu Ile Ser Thr Ser
Glu Ala Gln Ala Gly Phe 100 105
110 Lys Val Lys Leu Asp Phe Thr Gly Trp Arg Thr Val Gly Val Ser
Leu 115 120 125 Asn
Asn Asp Leu Glu Asn Arg Glu Met Thr Leu Asn Ala Thr Asn Thr 130
135 140 Ser Ser Asp Gly Thr Gln
Asp Ser Ile Gly Arg Ser Leu Gly Ala Lys 145 150
155 160 Val Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn
Val Ser Gln Gly Glu 165 170
175 Ile Tyr Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln
180 185 190 Trp Ser
Asp Tyr Gln Val Lys Thr Arg Leu Ser Glu Pro Glu Ile Gln 195
200 205 Phe His Asn Val Lys Pro Gln
Leu Pro Val Thr Pro Glu Asn Leu Ala 210 215
220 Ala Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu
Phe Val Gly Gly 225 230 235
240 Glu Lys Glu Thr Asn Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys
245 250 255 Ser Asp Phe
Asp Ala Leu Asn Thr His Thr Leu Ala Asn Gly Gly Thr 260
265 270 Gln Gly Arg His Leu Ile Thr Asp
Lys Gln Ile Ile Ile Tyr Gln Pro 275 280
285 Glu Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp Asn
Tyr Val Ile 290 295 300
Leu Gly Asn Tyr Thr Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val 305
310 315 320 Leu Glu Lys Asp
Pro Thr Gln Lys Ala Gln Leu Lys Gln Met Tyr Leu 325
330 335 Leu Met Thr Lys His Leu Leu Asp Gln
Gly Phe Val Lys Gly Ser Ala 340 345
350 Leu Val Thr Thr His His Trp Gly Tyr Ser Ser Arg Trp Trp
Tyr Ile 355 360 365
Ser Thr Leu Leu Met Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr 370
375 380 Gln Val Tyr Asp Ser
Leu Leu Trp Tyr Ser Arg Glu Phe Lys Ser Ser 385 390
395 400 Phe Asp Met Lys Val Ser Ala Asp Ser Ser
Asp Leu Asp Tyr Phe Asn 405 410
415 Thr Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu Pro Asp
Asp 420 425 430 Gln
Lys Arg Ile Asn Leu Val Asn Thr Phe Ser His Tyr Ile Thr Gly 435
440 445 Ala Leu Thr Gln Val Pro
Pro Gly Gly Lys Asp Gly Leu Arg Pro Asp 450 455
460 Gly Thr Ala Trp Arg His Glu Gly Asn Tyr Pro
Gly Tyr Ser Phe Pro 465 470 475
480 Ala Phe Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro
485 490 495 Phe Ser
Val Gly Glu Ser Gly Trp Asn Asn Leu Lys Lys Ala Met Val 500
505 510 Ser Ala Trp Ile Tyr Ser Asn
Pro Glu Val Gly Leu Pro Leu Ala Gly 515 520
525 Arg His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val
Ala Gln Gly Tyr 530 535 540
Tyr Trp Leu Ala Met Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala 545
550 555 560 Ser Ile Tyr
Leu Ala Ile Ser Asp Lys Thr Gln Asn Glu Ser Thr Ala 565
570 575 Ile Phe Gly Glu Thr Ile Thr Pro
Ala Ser Leu Pro Gln Gly Phe Tyr 580 585
590 Ala Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln
Asp Lys Met 595 600 605
Val Thr Leu Lys Ala Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr 610
615 620 Asn Lys Asp Asn
Arg Tyr Gly Arg Tyr Gln Ser His Gly Val Ala Gln 625 630
635 640 Ile Val Ser Asn Gly Ser Gln Leu Ser
Gln Gly Tyr Gln Gln Glu Gly 645 650
655 Trp Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile His Leu
Pro Leu 660 665 670
Lys Asp Leu Asp Ser Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu
675 680 685 Arg Gly Phe Ser
Gly Thr Ser Ser Leu Glu Gly Gln Tyr Gly Met Met 690
695 700 Ala Phe Asn Leu Ile Tyr Pro Ala
Asn Leu Glu Arg Phe Asp Pro Asn 705 710
715 720 Phe Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn
His Leu Ile Phe 725 730
735 Ile Gly Ser Asn Ile Asn Ser Ser Asp Lys Asn Lys Asn Val Glu Thr
740 745 750 Thr Leu Phe
Gln His Ala Ile Thr Pro Thr Leu Asn Thr Leu Trp Ile 755
760 765 Asn Gly Gln Lys Ile Glu Asn Met
Pro Tyr Gln Thr Thr Leu Gln Gln 770 775
780 Gly Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu
Ile Thr Gln 785 790 795
800 Ala Glu Lys Val Asn Val Ser Arg Gln His Gln Val Ser Ala Glu Asn
805 810 815 Lys Asn Arg Gln
Pro Thr Glu Gly Asn Phe Ser Ser Ala Trp Ile Asp 820
825 830 His Ser Thr Arg Pro Lys Asp Ala Ser
Tyr Glu Tyr Met Val Phe Leu 835 840
845 Asp Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys Phe
Arg Glu 850 855 860
Asn Asn Gly Leu Tyr Gln Val Leu Arg Lys Asp Lys Asp Val His Ile 865
870 875 880 Ile Leu Asp Lys Leu
Ser Asn Val Thr Gly Tyr Ala Phe Tyr Gln Pro 885
890 895 Ala Ser Ile Glu Asp Lys Trp Ile Lys Lys
Val Asn Lys Pro Ala 900 905
910 231637PRTArtificialSynthetic polypeptide, C terminal fusion
Chimera between ABCI-FL and GGF2-FL 23Ala Thr Ser Asn Pro Ala Phe Asp Pro
Lys Asn Leu Met Gln Ser Glu 1 5 10
15 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser
Ser Asp 20 25 30
Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn
35 40 45 Gln Ser Leu Leu
Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50
55 60 Lys Lys Leu Ile Val Pro Thr Asp
Lys Glu Ala Ser Lys Ala Trp Gly 65 70
75 80 Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr
Asn Glu Lys Pro 85 90
95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr
100 105 110 Ser Glu Ala
Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp 115
120 125 Arg Thr Val Gly Val Ser Leu Asn
Asn Asp Leu Glu Asn Arg Glu Met 130 135
140 Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln
Asp Ser Ile 145 150 155
160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro
165 170 175 Ser Asn Val Ser
Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser 180
185 190 Val Asp Asp Ala Arg Tyr Gln Trp Ser
Asp Tyr Gln Val Lys Thr Arg 195 200
205 Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln
Leu Pro 210 215 220
Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu 225
230 235 240 Ile Asn Glu Phe Val
Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu 245
250 255 Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe
Asp Ala Leu Asn Thr His 260 265
270 Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp
Lys 275 280 285 Gln
Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln 290
295 300 Leu Phe Asp Asn Tyr Val
Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305 310
315 320 Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp
Pro Thr Gln Lys Ala 325 330
335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln
340 345 350 Gly Phe
Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr 355
360 365 Ser Ser Arg Trp Trp Tyr Ile
Ser Thr Leu Leu Met Ser Asp Ala Leu 370 375
380 Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser
Leu Leu Trp Tyr 385 390 395
400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser
405 410 415 Ser Asp Leu
Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu 420
425 430 Leu Leu Leu Glu Pro Asp Asp Gln
Lys Arg Ile Asn Leu Val Asn Thr 435 440
445 Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro
Pro Gly Gly 450 455 460
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn 465
470 475 480 Tyr Pro Gly Tyr
Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile 485
490 495 Tyr Leu Leu Arg Asp Thr Pro Phe Ser
Val Gly Glu Ser Gly Trp Asn 500 505
510 Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn
Pro Glu 515 520 525
Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu 530
535 540 Lys Ser Val Ala Gln
Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545 550
555 560 Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr
Leu Ala Ile Ser Asp Lys 565 570
575 Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro
Ala 580 585 590 Ser
Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile 595
600 605 His Arg Trp Gln Asp Lys
Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610 615
620 Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn
Arg Tyr Gly Arg Tyr 625 630 635
640 Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser
645 650 655 Gln Gly
Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala 660
665 670 Thr Thr Ile His Leu Pro Leu
Lys Asp Leu Asp Ser Pro Lys Pro His 675 680
685 Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly
Thr Ser Ser Leu 690 695 700
Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn 705
710 715 720 Leu Glu Arg
Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala 725
730 735 Ala Asp Asn His Leu Ile Phe Ile
Gly Ser Asn Ile Asn Ser Ser Asp 740 745
750 Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His Ala
Ile Thr Pro 755 760 765
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro 770
775 780 Tyr Gln Thr Thr
Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785 790
795 800 Asn Gly Tyr Leu Ile Thr Gln Ala Glu
Lys Val Asn Val Ser Arg Gln 805 810
815 His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu
Gly Asn 820 825 830
Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser
835 840 845 Tyr Glu Tyr Met
Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850
855 860 Met Ala Gln Lys Phe Arg Glu Asn
Asn Gly Leu Tyr Gln Val Leu Arg 865 870
875 880 Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu
Ser Asn Val Thr 885 890
895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys
900 905 910 Lys Val Asn
Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr 915
920 925 Leu Ile Val Ser Ala Val Thr Pro
Asp Leu Asn Met Thr Arg Gln Lys 930 935
940 Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn Gly
Lys Trp Gln 945 950 955
960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn
965 970 975 Thr Glu Leu Thr
Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys 980
985 990 Leu Ser Pro Leu Pro Met Ser Glu
Arg Lys Glu Gly Arg Gly Lys Gly 995 1000
1005 Lys Gly Lys Lys Lys Glu Arg Gly Ser Gly Lys
Lys Pro Glu Ser 1010 1015 1020
Ala Ala Gly Ser Gln Ser Pro Ala Leu Pro Pro Arg Leu Lys Glu
1025 1030 1035 Met Lys Ser
Gln Glu Ser Ala Ala Gly Ser Lys Leu Val Leu Arg 1040
1045 1050 Cys Glu Thr Ser Ser Glu Tyr Ser
Ser Leu Arg Phe Lys Trp Phe 1055 1060
1065 Lys Asn Gly Asn Glu Leu Asn Arg Lys Asn Lys Pro Gln
Asn Ile 1070 1075 1080
Lys Ile Gln Lys Lys Pro Gly Lys Ser Glu Leu Arg Ile Asn Lys 1085
1090 1095 Ala Ser Leu Ala Asp
Ser Gly Glu Tyr Met Cys Lys Val Ile Ser 1100 1105
1110 Lys Leu Gly Asn Asp Ser Ala Ser Ala Asn
Ile Thr Ile Val Glu 1115 1120 1125
Ser Asn Glu Ile Ile Thr Gly Met Pro Ala Ser Thr Glu Gly Ala
1130 1135 1140 Tyr Val
Ser Ser Glu Ser Pro Ile Arg Ile Ser Val Ser Thr Glu 1145
1150 1155 Gly Ala Asn Thr Ser Ser Ser
Thr Ser Thr Ser Thr Thr Gly Thr 1160 1165
1170 Ser His Leu Val Lys Cys Ala Glu Lys Glu Lys Thr
Phe Cys Val 1175 1180 1185
Asn Gly Gly Glu Cys Phe Met Val Lys Asp Leu Ser Asn Pro Ser 1190
1195 1200 Arg Tyr Leu Cys Lys
Cys Gln Pro Gly Phe Thr Gly Ala Arg Cys 1205 1210
1215 Thr Glu Asn Val Pro Met Lys Val Gln Asn
Gln Glu Lys Ala Glu 1220 1225 1230
Glu Leu Tyr Gln Lys Arg Val Leu Thr Ile Thr Gly Ile Cys Ile
1235 1240 1245 Ala Leu
Leu Val Val Gly Ile Met Cys Val Val Ala Tyr Cys Lys 1250
1255 1260 Thr Lys Lys Gln Arg Lys Lys
Leu His Asp Arg Leu Arg Gln Ser 1265 1270
1275 Leu Arg Ser Glu Arg Asn Asn Met Met Asn Ile Ala
Asn Gly Pro 1280 1285 1290
His His Pro Asn Pro Pro Pro Glu Asn Val Gln Leu Val Asn Gln 1295
1300 1305 Tyr Val Ser Lys Asn
Val Ile Ser Ser Glu His Ile Val Glu Arg 1310 1315
1320 Glu Ala Glu Thr Ser Phe Ser Thr Ser His
Tyr Thr Ser Thr Ala 1325 1330 1335
His His Ser Thr Thr Val Thr Gln Thr Pro Ser His Ser Trp Ser
1340 1345 1350 Asn Gly
His Thr Glu Ser Ile Leu Ser Glu Ser His Ser Val Ile 1355
1360 1365 Val Met Ser Ser Val Glu Asn
Ser Arg His Ser Ser Pro Thr Gly 1370 1375
1380 Gly Pro Arg Gly Arg Leu Asn Gly Thr Gly Gly Pro
Arg Glu Cys 1385 1390 1395
Asn Ser Phe Leu Arg His Ala Arg Glu Thr Pro Asp Ser Tyr Arg 1400
1405 1410 Asp Ser Pro His Ser
Glu Arg Tyr Val Ser Ala Met Thr Thr Pro 1415 1420
1425 Ala Arg Met Ser Pro Val Asp Phe His Thr
Pro Ser Ser Pro Lys 1430 1435 1440
Ser Pro Pro Ser Glu Met Ser Pro Pro Val Ser Ser Met Thr Val
1445 1450 1455 Ser Met
Pro Ser Met Ala Val Ser Pro Phe Met Glu Glu Glu Arg 1460
1465 1470 Pro Leu Leu Leu Val Thr Pro
Pro Arg Leu Arg Glu Lys Lys Phe 1475 1480
1485 Asp His His Pro Gln Gln Phe Ser Ser Phe His His
Asn Pro Ala 1490 1495 1500
His Asp Ser Asn Ser Leu Pro Ala Ser Pro Leu Arg Ile Val Glu 1505
1510 1515 Asp Glu Glu Tyr Glu
Thr Thr Gln Glu Tyr Glu Pro Ala Gln Glu 1520 1525
1530 Pro Val Lys Lys Leu Ala Asn Ser Arg Arg
Ala Lys Arg Thr Lys 1535 1540 1545
Pro Asn Gly His Ile Ala Asn Arg Leu Glu Val Asp Ser Asn Thr
1550 1555 1560 Ser Ser
Gln Ser Ser Asn Ser Glu Ser Glu Thr Glu Asp Glu Arg 1565
1570 1575 Val Gly Glu Asp Thr Pro Phe
Leu Gly Ile Gln Asn Pro Leu Ala 1580 1585
1590 Ala Ser Leu Glu Ala Thr Pro Ala Phe Arg Leu Ala
Asp Ser Arg 1595 1600 1605
Thr Asn Pro Ala Gly Arg Phe Ser Thr Gln Glu Glu Ile Gln Ala 1610
1615 1620 Arg Leu Ser Ser Val
Ile Ala Asn Gln Asp Pro Ile Ala Val 1625 1630
1635 241150PRTArtificialSynthetic polypeptide, C
terminal fusion Chimera between chondroitinase ABCI-FL and
GGF2-L(sub)250-C(sub) 402 24Ala Thr Ser Asn Pro Ala Phe Asp Pro Lys
Asn Leu Met Gln Ser Glu 1 5 10
15 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser Ser
Asp 20 25 30 Lys
Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn 35
40 45 Gln Ser Leu Leu Trp Lys
Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50 55
60 Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala
Ser Lys Ala Trp Gly 65 70 75
80 Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys Pro
85 90 95 Ile Asp
Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr 100
105 110 Ser Glu Ala Gln Ala Gly Phe
Lys Val Lys Leu Asp Phe Thr Gly Trp 115 120
125 Arg Thr Val Gly Val Ser Leu Asn Asn Asp Leu Glu
Asn Arg Glu Met 130 135 140
Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile 145
150 155 160 Gly Arg Ser
Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro 165
170 175 Ser Asn Val Ser Gln Gly Glu Ile
Tyr Ile Asp Arg Ile Met Phe Ser 180 185
190 Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val
Lys Thr Arg 195 200 205
Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln Leu Pro 210
215 220 Val Thr Pro Glu
Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu 225 230
235 240 Ile Asn Glu Phe Val Gly Gly Glu Lys
Glu Thr Asn Leu Ala Leu Glu 245 250
255 Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn
Thr His 260 265 270
Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp Lys
275 280 285 Gln Ile Ile Ile
Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln 290
295 300 Leu Phe Asp Asn Tyr Val Ile Leu
Gly Asn Tyr Thr Thr Leu Met Phe 305 310
315 320 Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro
Thr Gln Lys Ala 325 330
335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln
340 345 350 Gly Phe Val
Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr 355
360 365 Ser Ser Arg Trp Trp Tyr Ile Ser
Thr Leu Leu Met Ser Asp Ala Leu 370 375
380 Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser Leu
Leu Trp Tyr 385 390 395
400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser
405 410 415 Ser Asp Leu Asp
Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu 420
425 430 Leu Leu Leu Glu Pro Asp Asp Gln Lys
Arg Ile Asn Leu Val Asn Thr 435 440
445 Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro
Gly Gly 450 455 460
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn 465
470 475 480 Tyr Pro Gly Tyr Ser
Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile 485
490 495 Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val
Gly Glu Ser Gly Trp Asn 500 505
510 Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn Pro
Glu 515 520 525 Val
Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu 530
535 540 Lys Ser Val Ala Gln Gly
Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545 550
555 560 Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu
Ala Ile Ser Asp Lys 565 570
575 Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro Ala
580 585 590 Ser Leu
Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile 595
600 605 His Arg Trp Gln Asp Lys Met
Val Thr Leu Lys Ala Tyr Asn Thr Asn 610 615
620 Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg
Tyr Gly Arg Tyr 625 630 635
640 Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser
645 650 655 Gln Gly Tyr
Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala 660
665 670 Thr Thr Ile His Leu Pro Leu Lys
Asp Leu Asp Ser Pro Lys Pro His 675 680
685 Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr
Ser Ser Leu 690 695 700
Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn 705
710 715 720 Leu Glu Arg Phe
Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala 725
730 735 Ala Asp Asn His Leu Ile Phe Ile Gly
Ser Asn Ile Asn Ser Ser Asp 740 745
750 Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His Ala Ile
Thr Pro 755 760 765
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro 770
775 780 Tyr Gln Thr Thr Leu
Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785 790
795 800 Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys
Val Asn Val Ser Arg Gln 805 810
815 His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu Gly
Asn 820 825 830 Phe
Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser 835
840 845 Tyr Glu Tyr Met Val Phe
Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850 855
860 Met Ala Gln Lys Phe Arg Glu Asn Asn Gly Leu
Tyr Gln Val Leu Arg 865 870 875
880 Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu Ser Asn Val Thr
885 890 895 Gly Tyr
Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys 900
905 910 Lys Val Asn Lys Pro Ala Ile
Val Met Thr His Arg Gln Lys Asp Thr 915 920
925 Leu Ile Val Ser Ala Val Thr Pro Asp Leu Asn Met
Thr Arg Gln Lys 930 935 940
Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn Gly Lys Trp Gln 945
950 955 960 Ser Ala Asp
Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn 965
970 975 Thr Glu Leu Thr Phe Thr Ser Tyr
Phe Gly Ile Pro Gln Glu Ile Lys 980 985
990 Leu Ser Pro Leu Pro Cys Ile Ala Leu Leu Val Val
Gly Ile Met Cys 995 1000 1005
Val Val Ala Tyr Cys Lys Thr Lys Lys Gln Arg Lys Lys Leu His
1010 1015 1020 Asp Arg Leu
Arg Gln Ser Leu Arg Ser Glu Arg Asn Asn Met Met 1025
1030 1035 Asn Ile Ala Asn Gly Pro His His
Pro Asn Pro Pro Pro Glu Asn 1040 1045
1050 Val Gln Leu Val Asn Gln Tyr Val Ser Lys Asn Val Ile
Ser Ser 1055 1060 1065
Glu His Ile Val Glu Arg Glu Ala Glu Thr Ser Phe Ser Thr Ser 1070
1075 1080 His Tyr Thr Ser Thr
Ala His His Ser Thr Thr Val Thr Gln Thr 1085 1090
1095 Pro Ser His Ser Trp Ser Asn Gly His Thr
Glu Ser Ile Leu Ser 1100 1105 1110
Glu Ser His Ser Val Ile Val Met Ser Ser Val Glu Asn Ser Arg
1115 1120 1125 His Ser
Ser Pro Thr Gly Gly Pro Arg Gly Arg Leu Asn Gly Thr 1130
1135 1140 Gly Gly Pro Arg Glu Cys Asn
1145 1150 251170PRTArtificialSynthetic polypeptide, C
terminal fusion Chimera between ABCI-FL and GGF2-L(sub)250-E(sub)422
25Ala Thr Ser Asn Pro Ala Phe Asp Pro Lys Asn Leu Met Gln Ser Glu 1
5 10 15 Ile Tyr His Phe
Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser Ser Asp 20
25 30 Lys Asn Ser Ile Leu Thr Leu Ser Asp
Lys Arg Ser Ile Met Gly Asn 35 40
45 Gln Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr
Leu His 50 55 60
Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly 65
70 75 80 Arg Ser Ser Thr Pro
Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys Pro 85
90 95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly
Glu Lys Leu Ile Ser Thr 100 105
110 Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly
Trp 115 120 125 Arg
Thr Val Gly Val Ser Leu Asn Asn Asp Leu Glu Asn Arg Glu Met 130
135 140 Thr Leu Asn Ala Thr Asn
Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile 145 150
155 160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile
Arg Phe Lys Ala Pro 165 170
175 Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser
180 185 190 Val Asp
Asp Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val Lys Thr Arg 195
200 205 Leu Ser Glu Pro Glu Ile Gln
Phe His Asn Val Lys Pro Gln Leu Pro 210 215
220 Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile
Arg Gln Arg Leu 225 230 235
240 Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu
245 250 255 Glu Asn Ile
Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr His 260
265 270 Thr Leu Ala Asn Gly Gly Thr Gln
Gly Arg His Leu Ile Thr Asp Lys 275 280
285 Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln
Asp Lys Gln 290 295 300
Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305
310 315 320 Asn Ile Ser Arg
Ala Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys Ala 325
330 335 Gln Leu Lys Gln Met Tyr Leu Leu Met
Thr Lys His Leu Leu Asp Gln 340 345
350 Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp
Gly Tyr 355 360 365
Ser Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser Asp Ala Leu 370
375 380 Lys Glu Ala Asn Leu
Gln Thr Gln Val Tyr Asp Ser Leu Leu Trp Tyr 385 390
395 400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met
Lys Val Ser Ala Asp Ser 405 410
415 Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala
Leu 420 425 430 Leu
Leu Leu Glu Pro Asp Asp Gln Lys Arg Ile Asn Leu Val Asn Thr 435
440 445 Phe Ser His Tyr Ile Thr
Gly Ala Leu Thr Gln Val Pro Pro Gly Gly 450 455
460 Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp
Arg His Glu Gly Asn 465 470 475
480 Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile
485 490 495 Tyr Leu
Leu Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly Trp Asn 500
505 510 Asn Leu Lys Lys Ala Met Val
Ser Ala Trp Ile Tyr Ser Asn Pro Glu 515 520
525 Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn
Ser Pro Ser Leu 530 535 540
Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545
550 555 560 Ser Pro Asp
Lys Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser Asp Lys 565
570 575 Thr Gln Asn Glu Ser Thr Ala Ile
Phe Gly Glu Thr Ile Thr Pro Ala 580 585
590 Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala
Phe Gly Ile 595 600 605
His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610
615 620 Val Trp Ser Ser
Glu Ile Tyr Asn Lys Asp Asn Arg Tyr Gly Arg Tyr 625 630
635 640 Gln Ser His Gly Val Ala Gln Ile Val
Ser Asn Gly Ser Gln Leu Ser 645 650
655 Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu
Gly Ala 660 665 670
Thr Thr Ile His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His
675 680 685 Thr Leu Met Gln
Arg Gly Glu Arg Gly Phe Ser Gly Thr Ser Ser Leu 690
695 700 Glu Gly Gln Tyr Gly Met Met Ala
Phe Asn Leu Ile Tyr Pro Ala Asn 705 710
715 720 Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys
Ser Val Leu Ala 725 730
735 Ala Asp Asn His Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp
740 745 750 Lys Asn Lys
Asn Val Glu Thr Thr Leu Phe Gln His Ala Ile Thr Pro 755
760 765 Thr Leu Asn Thr Leu Trp Ile Asn
Gly Gln Lys Ile Glu Asn Met Pro 770 775
780 Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp
Ser Asn Gly 785 790 795
800 Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln
805 810 815 His Gln Val Ser
Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu Gly Asn 820
825 830 Phe Ser Ser Ala Trp Ile Asp His Ser
Thr Arg Pro Lys Asp Ala Ser 835 840
845 Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys Met
Gly Glu 850 855 860
Met Ala Gln Lys Phe Arg Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg 865
870 875 880 Lys Asp Lys Asp Val
His Ile Ile Leu Asp Lys Leu Ser Asn Val Thr 885
890 895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile
Glu Asp Lys Trp Ile Lys 900 905
910 Lys Val Asn Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp
Thr 915 920 925 Leu
Ile Val Ser Ala Val Thr Pro Asp Leu Asn Met Thr Arg Gln Lys 930
935 940 Ala Ala Thr Pro Val Thr
Ile Asn Val Thr Ile Asn Gly Lys Trp Gln 945 950
955 960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln
Val Ser Gly Asp Asn 965 970
975 Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys
980 985 990 Leu Ser
Pro Leu Pro Cys Ile Ala Leu Leu Val Val Gly Ile Met Cys 995
1000 1005 Val Val Ala Tyr Cys
Lys Thr Lys Lys Gln Arg Lys Lys Leu His 1010 1015
1020 Asp Arg Leu Arg Gln Ser Leu Arg Ser Glu
Arg Asn Asn Met Met 1025 1030 1035
Asn Ile Ala Asn Gly Pro His His Pro Asn Pro Pro Pro Glu Asn
1040 1045 1050 Val Gln
Leu Val Asn Gln Tyr Val Ser Lys Asn Val Ile Ser Ser 1055
1060 1065 Glu His Ile Val Glu Arg Glu
Ala Glu Thr Ser Phe Ser Thr Ser 1070 1075
1080 His Tyr Thr Ser Thr Ala His His Ser Thr Thr Val
Thr Gln Thr 1085 1090 1095
Pro Ser His Ser Trp Ser Asn Gly His Thr Glu Ser Ile Leu Ser 1100
1105 1110 Glu Ser His Ser Val
Ile Val Met Ser Ser Val Glu Asn Ser Arg 1115 1120
1125 His Ser Ser Pro Thr Gly Gly Pro Arg Gly
Arg Leu Asn Gly Thr 1130 1135 1140
Gly Gly Pro Arg Glu Cys Asn Ser Phe Leu Arg His Ala Arg Glu
1145 1150 1155 Thr Pro
Asp Ser Tyr Arg Asp Ser Pro His Ser Glu 1160 1165
1170 261050PRTArtificialSynthetic polypeptide, C terminal
fusion Chimera between chondroitinase ABCI-FL and
GGF2-T(sub)350-C(sub) 402 26Ala Thr Ser Asn Pro Ala Phe Asp Pro Lys
Asn Leu Met Gln Ser Glu 1 5 10
15 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser Ser
Asp 20 25 30 Lys
Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn 35
40 45 Gln Ser Leu Leu Trp Lys
Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50 55
60 Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala
Ser Lys Ala Trp Gly 65 70 75
80 Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys Pro
85 90 95 Ile Asp
Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr 100
105 110 Ser Glu Ala Gln Ala Gly Phe
Lys Val Lys Leu Asp Phe Thr Gly Trp 115 120
125 Arg Thr Val Gly Val Ser Leu Asn Asn Asp Leu Glu
Asn Arg Glu Met 130 135 140
Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile 145
150 155 160 Gly Arg Ser
Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro 165
170 175 Ser Asn Val Ser Gln Gly Glu Ile
Tyr Ile Asp Arg Ile Met Phe Ser 180 185
190 Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val
Lys Thr Arg 195 200 205
Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln Leu Pro 210
215 220 Val Thr Pro Glu
Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu 225 230
235 240 Ile Asn Glu Phe Val Gly Gly Glu Lys
Glu Thr Asn Leu Ala Leu Glu 245 250
255 Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn
Thr His 260 265 270
Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp Lys
275 280 285 Gln Ile Ile Ile
Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln 290
295 300 Leu Phe Asp Asn Tyr Val Ile Leu
Gly Asn Tyr Thr Thr Leu Met Phe 305 310
315 320 Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro
Thr Gln Lys Ala 325 330
335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln
340 345 350 Gly Phe Val
Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr 355
360 365 Ser Ser Arg Trp Trp Tyr Ile Ser
Thr Leu Leu Met Ser Asp Ala Leu 370 375
380 Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser Leu
Leu Trp Tyr 385 390 395
400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser
405 410 415 Ser Asp Leu Asp
Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu 420
425 430 Leu Leu Leu Glu Pro Asp Asp Gln Lys
Arg Ile Asn Leu Val Asn Thr 435 440
445 Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro
Gly Gly 450 455 460
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn 465
470 475 480 Tyr Pro Gly Tyr Ser
Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile 485
490 495 Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val
Gly Glu Ser Gly Trp Asn 500 505
510 Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn Pro
Glu 515 520 525 Val
Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu 530
535 540 Lys Ser Val Ala Gln Gly
Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545 550
555 560 Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu
Ala Ile Ser Asp Lys 565 570
575 Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro Ala
580 585 590 Ser Leu
Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile 595
600 605 His Arg Trp Gln Asp Lys Met
Val Thr Leu Lys Ala Tyr Asn Thr Asn 610 615
620 Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg
Tyr Gly Arg Tyr 625 630 635
640 Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser
645 650 655 Gln Gly Tyr
Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala 660
665 670 Thr Thr Ile His Leu Pro Leu Lys
Asp Leu Asp Ser Pro Lys Pro His 675 680
685 Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr
Ser Ser Leu 690 695 700
Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn 705
710 715 720 Leu Glu Arg Phe
Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala 725
730 735 Ala Asp Asn His Leu Ile Phe Ile Gly
Ser Asn Ile Asn Ser Ser Asp 740 745
750 Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His Ala Ile
Thr Pro 755 760 765
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro 770
775 780 Tyr Gln Thr Thr Leu
Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785 790
795 800 Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys
Val Asn Val Ser Arg Gln 805 810
815 His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu Gly
Asn 820 825 830 Phe
Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser 835
840 845 Tyr Glu Tyr Met Val Phe
Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850 855
860 Met Ala Gln Lys Phe Arg Glu Asn Asn Gly Leu
Tyr Gln Val Leu Arg 865 870 875
880 Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu Ser Asn Val Thr
885 890 895 Gly Tyr
Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys 900
905 910 Lys Val Asn Lys Pro Ala Ile
Val Met Thr His Arg Gln Lys Asp Thr 915 920
925 Leu Ile Val Ser Ala Val Thr Pro Asp Leu Asn Met
Thr Arg Gln Lys 930 935 940
Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn Gly Lys Trp Gln 945
950 955 960 Ser Ala Asp
Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn 965
970 975 Thr Glu Leu Thr Phe Thr Ser Tyr
Phe Gly Ile Pro Gln Glu Ile Lys 980 985
990 Leu Ser Pro Leu Pro Thr Pro Ser His Ser Trp Ser
Asn Gly His Thr 995 1000 1005
Glu Ser Ile Leu Ser Glu Ser His Ser Val Ile Val Met Ser Ser
1010 1015 1020 Val Glu Asn
Ser Arg His Ser Ser Pro Thr Gly Gly Pro Arg Gly 1025
1030 1035 Arg Leu Asn Gly Thr Gly Gly Pro
Arg Glu Cys Asn 1040 1045 1050
271281PRTArtificialSynthetic polypeptide, N terminal fusion chimera
between NgR(sub)27-311 and chondroitinase ABCI without a peptide
spacer or linking group 27Cys Pro Gly Ala Cys Val Cys Tyr Asn Glu Pro Lys
Val Thr Thr Ser 1 5 10
15 Cys Pro Gln Gln Gly Leu Gln Ala Val Pro Val Gly Ile Pro Ala Ala
20 25 30 Ser Gln Arg
Ile Phe Leu His Gly Asn Arg Ile Ser His Val Pro Ala 35
40 45 Ala Ser Phe Arg Ala Cys Arg Asn
Leu Thr Ile Leu Trp Leu His Ser 50 55
60 Asn Val Leu Ala Arg Ile Asp Ala Ala Ala Phe Thr Gly
Leu Ala Leu 65 70 75
80 Leu Glu Gln Leu Asp Leu Ser Asp Asn Ala Gln Leu Arg Ser Val Asp
85 90 95 Pro Ala Thr Phe
His Gly Leu Gly Arg Leu His Thr Leu His Leu Asp 100
105 110 Arg Cys Gly Leu Gln Glu Leu Gly Pro
Gly Leu Phe Arg Gly Leu Ala 115 120
125 Ala Leu Gln Tyr Leu Tyr Leu Gln Asp Asn Ala Leu Gln Ala
Leu Pro 130 135 140
Asp Asp Thr Phe Arg Asp Leu Gly Asn Leu Thr His Leu Phe Leu His 145
150 155 160 Gly Asn Arg Ile Ser
Ser Val Pro Glu Arg Ala Phe Arg Gly Leu His 165
170 175 Ser Leu Asp Arg Leu Leu Leu His Gln Asn
Arg Val Ala His Val His 180 185
190 Pro His Ala Phe Arg Asp Leu Gly Arg Leu Met Thr Leu Tyr Leu
Phe 195 200 205 Ala
Asn Asn Leu Ser Ala Leu Pro Thr Glu Ala Leu Ala Pro Leu Arg 210
215 220 Ala Leu Gln Tyr Leu Arg
Leu Asn Asp Asn Pro Trp Val Cys Asp Cys 225 230
235 240 Arg Ala Arg Pro Leu Trp Ala Trp Leu Gln Lys
Phe Arg Gly Ser Ser 245 250
255 Ser Glu Val Pro Cys Ser Leu Pro Gln Arg Leu Ala Gly Arg Asp Leu
260 265 270 Lys Arg
Leu Ala Ala Asn Asp Leu Gln Gly Cys Ala Val Ala Thr Ser 275
280 285 Asn Pro Ala Phe Asp Pro Lys
Asn Leu Met Gln Ser Glu Ile Tyr His 290 295
300 Phe Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser Ser
Asp Lys Asn Ser 305 310 315
320 Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn Gln Ser Leu
325 330 335 Leu Trp Lys
Trp Lys Gly Gly Ser Ser Phe Thr Leu His Lys Lys Leu 340
345 350 Ile Val Pro Thr Asp Lys Glu Ala
Ser Lys Ala Trp Gly Arg Ser Ser 355 360
365 Thr Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys Pro
Ile Asp Gly 370 375 380
Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala 385
390 395 400 Gln Ala Gly Phe
Lys Val Lys Leu Asp Phe Thr Gly Trp Arg Thr Val 405
410 415 Gly Val Ser Leu Asn Asn Asp Leu Glu
Asn Arg Glu Met Thr Leu Asn 420 425
430 Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile Gly
Arg Ser 435 440 445
Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn Val 450
455 460 Ser Gln Gly Glu Ile
Tyr Ile Asp Arg Ile Met Phe Ser Val Asp Asp 465 470
475 480 Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val
Lys Thr Arg Leu Ser Glu 485 490
495 Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln Leu Pro Val Thr
Pro 500 505 510 Glu
Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu 515
520 525 Phe Val Gly Gly Glu Lys
Glu Thr Asn Leu Ala Leu Glu Glu Asn Ile 530 535
540 Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn
Thr His Thr Leu Ala 545 550 555
560 Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp Lys Gln Ile Ile
565 570 575 Ile Tyr
Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp 580
585 590 Asn Tyr Val Ile Leu Gly Asn
Tyr Thr Thr Leu Met Phe Asn Ile Ser 595 600
605 Arg Ala Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys
Ala Gln Leu Lys 610 615 620
Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln Gly Phe Val 625
630 635 640 Lys Gly Ser
Ala Leu Val Thr Thr His His Trp Gly Tyr Ser Ser Arg 645
650 655 Trp Trp Tyr Ile Ser Thr Leu Leu
Met Ser Asp Ala Leu Lys Glu Ala 660 665
670 Asn Leu Gln Thr Gln Val Tyr Asp Ser Leu Leu Trp Tyr
Ser Arg Glu 675 680 685
Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser Ser Asp Leu 690
695 700 Asp Tyr Phe Asn
Thr Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu 705 710
715 720 Glu Pro Asp Asp Gln Lys Arg Ile Asn
Leu Val Asn Thr Phe Ser His 725 730
735 Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro Gly Gly Lys
Asp Gly 740 745 750
Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn Tyr Pro Gly
755 760 765 Tyr Ser Phe Pro
Ala Phe Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu 770
775 780 Arg Asp Thr Pro Phe Ser Val Gly
Glu Ser Gly Trp Asn Asn Leu Lys 785 790
795 800 Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn Pro
Glu Val Gly Leu 805 810
815 Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val
820 825 830 Ala Gln Gly
Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser Ser Pro Asp 835
840 845 Lys Thr Leu Ala Ser Ile Tyr Leu
Ala Ile Ser Asp Lys Thr Gln Asn 850 855
860 Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro Ala
Ser Leu Pro 865 870 875
880 Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp
885 890 895 Gln Asp Lys Met
Val Thr Leu Lys Ala Tyr Asn Thr Asn Val Trp Ser 900
905 910 Ser Glu Ile Tyr Asn Lys Asp Asn Arg
Tyr Gly Arg Tyr Gln Ser His 915 920
925 Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser Gln
Gly Tyr 930 935 940
Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile 945
950 955 960 His Leu Pro Leu Lys
Asp Leu Asp Ser Pro Lys Pro His Thr Leu Met 965
970 975 Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr
Ser Ser Leu Glu Gly Gln 980 985
990 Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn Leu
Glu Arg 995 1000 1005
Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala Ala Asp 1010
1015 1020 Asn His Leu Ile Phe
Ile Gly Ser Asn Ile Asn Ser Ser Asp Lys 1025 1030
1035 Asn Lys Asn Val Glu Thr Thr Leu Phe Gln
His Ala Ile Thr Pro 1040 1045 1050
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met
1055 1060 1065 Pro Tyr
Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser 1070
1075 1080 Asn Gly Asn Gly Tyr Leu Ile
Thr Gln Ala Glu Lys Val Asn Val 1085 1090
1095 Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys Asn
Arg Gln Pro 1100 1105 1110
Thr Glu Gly Asn Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg 1115
1120 1125 Pro Lys Asp Ala Ser
Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr 1130 1135
1140 Pro Glu Lys Met Gly Glu Met Ala Gln Lys
Phe Arg Glu Asn Asn 1145 1150 1155
Gly Leu Tyr Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile
1160 1165 1170 Leu Asp
Lys Leu Ser Asn Val Thr Gly Tyr Ala Phe Tyr Gln Pro 1175
1180 1185 Ala Ser Ile Glu Asp Lys Trp
Ile Lys Lys Val Asn Lys Pro Ala 1190 1195
1200 Ile Val Met Thr His Arg Gln Lys Asp Thr Leu Ile
Val Ser Ala 1205 1210 1215
Val Thr Pro Asp Leu Asn Met Thr Arg Gln Lys Ala Ala Thr Pro 1220
1225 1230 Val Thr Ile Asn Val
Thr Ile Asn Gly Lys Trp Gln Ser Ala Asp 1235 1240
1245 Lys Asn Ser Glu Val Lys Tyr Gln Val Ser
Gly Asp Asn Thr Glu 1250 1255 1260
Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys Leu
1265 1270 1275 Ser Pro
Leu 1280 281287PRTArtificialSynthetic polypeptide, N terminal
fusion chimera between NgR(sub)27-311 and chondoitinase ABCI with a
peptide spacer or linking group 28Cys Pro Gly Ala Cys Val Cys Tyr Asn
Glu Pro Lys Val Thr Thr Ser 1 5 10
15 Cys Pro Gln Gln Gly Leu Gln Ala Val Pro Val Gly Ile Pro
Ala Ala 20 25 30
Ser Gln Arg Ile Phe Leu His Gly Asn Arg Ile Ser His Val Pro Ala
35 40 45 Ala Ser Phe Arg
Ala Cys Arg Asn Leu Thr Ile Leu Trp Leu His Ser 50
55 60 Asn Val Leu Ala Arg Ile Asp Ala
Ala Ala Phe Thr Gly Leu Ala Leu 65 70
75 80 Leu Glu Gln Leu Asp Leu Ser Asp Asn Ala Gln Leu
Arg Ser Val Asp 85 90
95 Pro Ala Thr Phe His Gly Leu Gly Arg Leu His Thr Leu His Leu Asp
100 105 110 Arg Cys Gly
Leu Gln Glu Leu Gly Pro Gly Leu Phe Arg Gly Leu Ala 115
120 125 Ala Leu Gln Tyr Leu Tyr Leu Gln
Asp Asn Ala Leu Gln Ala Leu Pro 130 135
140 Asp Asp Thr Phe Arg Asp Leu Gly Asn Leu Thr His Leu
Phe Leu His 145 150 155
160 Gly Asn Arg Ile Ser Ser Val Pro Glu Arg Ala Phe Arg Gly Leu His
165 170 175 Ser Leu Asp Arg
Leu Leu Leu His Gln Asn Arg Val Ala His Val His 180
185 190 Pro His Ala Phe Arg Asp Leu Gly Arg
Leu Met Thr Leu Tyr Leu Phe 195 200
205 Ala Asn Asn Leu Ser Ala Leu Pro Thr Glu Ala Leu Ala Pro
Leu Arg 210 215 220
Ala Leu Gln Tyr Leu Arg Leu Asn Asp Asn Pro Trp Val Cys Asp Cys 225
230 235 240 Arg Ala Arg Pro Leu
Trp Ala Trp Leu Gln Lys Phe Arg Gly Ser Ser 245
250 255 Ser Glu Val Pro Cys Ser Leu Pro Gln Arg
Leu Ala Gly Arg Asp Leu 260 265
270 Lys Arg Leu Ala Ala Asn Asp Leu Gln Gly Cys Ala Val Gly Gly
Gly 275 280 285 Gly
Gly Ala Thr Ser Asn Pro Ala Phe Asp Pro Lys Asn Leu Met Gln 290
295 300 Ser Glu Ile Tyr His Phe
Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser 305 310
315 320 Ser Asp Lys Asn Ser Ile Leu Thr Leu Ser Asp
Lys Arg Ser Ile Met 325 330
335 Gly Asn Gln Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr
340 345 350 Leu His
Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala Ser Lys Ala 355
360 365 Trp Gly Arg Ser Ser Thr Pro
Val Phe Ser Phe Trp Leu Tyr Asn Glu 370 375
380 Lys Pro Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly
Glu Lys Leu Ile 385 390 395
400 Ser Thr Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr
405 410 415 Gly Trp Arg
Thr Val Gly Val Ser Leu Asn Asn Asp Leu Glu Asn Arg 420
425 430 Glu Met Thr Leu Asn Ala Thr Asn
Thr Ser Ser Asp Gly Thr Gln Asp 435 440
445 Ser Ile Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile
Arg Phe Lys 450 455 460
Ala Pro Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met 465
470 475 480 Phe Ser Val Asp
Asp Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val Lys 485
490 495 Thr Arg Leu Ser Glu Pro Glu Ile Gln
Phe His Asn Val Lys Pro Gln 500 505
510 Leu Pro Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile
Arg Gln 515 520 525
Arg Leu Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn Leu Ala 530
535 540 Leu Glu Glu Asn Ile
Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn 545 550
555 560 Thr His Thr Leu Ala Asn Gly Gly Thr Gln
Gly Arg His Leu Ile Thr 565 570
575 Asp Lys Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln
Asp 580 585 590 Lys
Gln Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu 595
600 605 Met Phe Asn Ile Ser Arg
Ala Tyr Val Leu Glu Lys Asp Pro Thr Gln 610 615
620 Lys Ala Gln Leu Lys Gln Met Tyr Leu Leu Met
Thr Lys His Leu Leu 625 630 635
640 Asp Gln Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp
645 650 655 Gly Tyr
Ser Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser Asp 660
665 670 Ala Leu Lys Glu Ala Asn Leu
Gln Thr Gln Val Tyr Asp Ser Leu Leu 675 680
685 Trp Tyr Ser Arg Glu Phe Lys Ser Ser Phe Asp Met
Lys Val Ser Ala 690 695 700
Asp Ser Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu 705
710 715 720 Ala Leu Leu
Leu Leu Glu Pro Asp Asp Gln Lys Arg Ile Asn Leu Val 725
730 735 Asn Thr Phe Ser His Tyr Ile Thr
Gly Ala Leu Thr Gln Val Pro Pro 740 745
750 Gly Gly Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp
Arg His Glu 755 760 765
Gly Asn Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln 770
775 780 Leu Ile Tyr Leu
Leu Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly 785 790
795 800 Trp Asn Asn Leu Lys Lys Ala Met Val
Ser Ala Trp Ile Tyr Ser Asn 805 810
815 Pro Glu Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn
Ser Pro 820 825 830
Ser Leu Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala
835 840 845 Lys Ser Ser Pro
Asp Lys Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser 850
855 860 Asp Lys Thr Gln Asn Glu Ser Thr
Ala Ile Phe Gly Glu Thr Ile Thr 865 870
875 880 Pro Ala Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn
Gly Gly Ala Phe 885 890
895 Gly Ile His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala Tyr Asn
900 905 910 Thr Asn Val
Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg Tyr Gly 915
920 925 Arg Tyr Gln Ser His Gly Val Ala
Gln Ile Val Ser Asn Gly Ser Gln 930 935
940 Leu Ser Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn
Arg Met Glu 945 950 955
960 Gly Ala Thr Thr Ile His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys
965 970 975 Pro His Thr Leu
Met Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr Ser 980
985 990 Ser Leu Glu Gly Gln Tyr Gly Met
Met Ala Phe Asn Leu Ile Tyr Pro 995 1000
1005 Ala Asn Leu Glu Arg Phe Asp Pro Asn Phe Thr
Ala Lys Lys Ser 1010 1015 1020
Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile Gly Ser Asn Ile
1025 1030 1035 Asn Ser Ser
Asp Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln 1040
1045 1050 His Ala Ile Thr Pro Thr Leu Asn
Thr Leu Trp Ile Asn Gly Gln 1055 1060
1065 Lys Ile Glu Asn Met Pro Tyr Gln Thr Thr Leu Gln Gln
Gly Asp 1070 1075 1080
Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala 1085
1090 1095 Glu Lys Val Asn Val
Ser Arg Gln His Gln Val Ser Ala Glu Asn 1100 1105
1110 Lys Asn Arg Gln Pro Thr Glu Gly Asn Phe
Ser Ser Ala Trp Ile 1115 1120 1125
Asp His Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr Met Val
1130 1135 1140 Phe Leu
Asp Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys 1145
1150 1155 Phe Arg Glu Asn Asn Gly Leu
Tyr Gln Val Leu Arg Lys Asp Lys 1160 1165
1170 Asp Val His Ile Ile Leu Asp Lys Leu Ser Asn Val
Thr Gly Tyr 1175 1180 1185
Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys Lys 1190
1195 1200 Val Asn Lys Pro Ala
Ile Val Met Thr His Arg Gln Lys Asp Thr 1205 1210
1215 Leu Ile Val Ser Ala Val Thr Pro Asp Leu
Asn Met Thr Arg Gln 1220 1225 1230
Lys Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn Gly Lys
1235 1240 1245 Trp Gln
Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser 1250
1255 1260 Gly Asp Asn Thr Glu Leu Thr
Phe Thr Ser Tyr Phe Gly Ile Pro 1265 1270
1275 Gln Glu Ile Lys Leu Ser Pro Leu Pro 1280
1285 291282PRTArtificialSynthetic Polypeptide, C
terminal fusion chimera between NgR(sub)27-311 and chondroitinase
ABCI protein without a peptide spacer or linking group 29Ala Thr Ser
Asn Pro Ala Phe Asp Pro Lys Asn Leu Met Gln Ser Glu 1 5
10 15 Ile Tyr His Phe Ala Gln Asn Asn
Pro Leu Ala Asp Phe Ser Ser Asp 20 25
30 Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile
Met Gly Asn 35 40 45
Gln Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50
55 60 Lys Lys Leu Ile
Val Pro Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly 65 70
75 80 Arg Ser Ser Thr Pro Val Phe Ser Phe
Trp Leu Tyr Asn Glu Lys Pro 85 90
95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile
Ser Thr 100 105 110
Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp
115 120 125 Arg Thr Val Gly
Val Ser Leu Asn Asn Asp Leu Glu Asn Arg Glu Met 130
135 140 Thr Leu Asn Ala Thr Asn Thr Ser
Ser Asp Gly Thr Gln Asp Ser Ile 145 150
155 160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg
Phe Lys Ala Pro 165 170
175 Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser
180 185 190 Val Asp Asp
Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val Lys Thr Arg 195
200 205 Leu Ser Glu Pro Glu Ile Gln Phe
His Asn Val Lys Pro Gln Leu Pro 210 215
220 Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg
Gln Arg Leu 225 230 235
240 Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu
245 250 255 Glu Asn Ile Ser
Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr His 260
265 270 Thr Leu Ala Asn Gly Gly Thr Gln Gly
Arg His Leu Ile Thr Asp Lys 275 280
285 Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp
Lys Gln 290 295 300
Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305
310 315 320 Asn Ile Ser Arg Ala
Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys Ala 325
330 335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr
Lys His Leu Leu Asp Gln 340 345
350 Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly
Tyr 355 360 365 Ser
Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser Asp Ala Leu 370
375 380 Lys Glu Ala Asn Leu Gln
Thr Gln Val Tyr Asp Ser Leu Leu Trp Tyr 385 390
395 400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys
Val Ser Ala Asp Ser 405 410
415 Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu
420 425 430 Leu Leu
Leu Glu Pro Asp Asp Gln Lys Arg Ile Asn Leu Val Asn Thr 435
440 445 Phe Ser His Tyr Ile Thr Gly
Ala Leu Thr Gln Val Pro Pro Gly Gly 450 455
460 Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg
His Glu Gly Asn 465 470 475
480 Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile
485 490 495 Tyr Leu Leu
Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly Trp Asn 500
505 510 Asn Leu Lys Lys Ala Met Val Ser
Ala Trp Ile Tyr Ser Asn Pro Glu 515 520
525 Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser
Pro Ser Leu 530 535 540
Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545
550 555 560 Ser Pro Asp Lys
Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser Asp Lys 565
570 575 Thr Gln Asn Glu Ser Thr Ala Ile Phe
Gly Glu Thr Ile Thr Pro Ala 580 585
590 Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe
Gly Ile 595 600 605
His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610
615 620 Val Trp Ser Ser Glu
Ile Tyr Asn Lys Asp Asn Arg Tyr Gly Arg Tyr 625 630
635 640 Gln Ser His Gly Val Ala Gln Ile Val Ser
Asn Gly Ser Gln Leu Ser 645 650
655 Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly
Ala 660 665 670 Thr
Thr Ile His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His 675
680 685 Thr Leu Met Gln Arg Gly
Glu Arg Gly Phe Ser Gly Thr Ser Ser Leu 690 695
700 Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu
Ile Tyr Pro Ala Asn 705 710 715
720 Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala
725 730 735 Ala Asp
Asn His Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp 740
745 750 Lys Asn Lys Asn Val Glu Thr
Thr Leu Phe Gln His Ala Ile Thr Pro 755 760
765 Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile
Glu Asn Met Pro 770 775 780
Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785
790 795 800 Asn Gly Tyr
Leu Ile Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln 805
810 815 His Gln Val Ser Ala Glu Asn Lys
Asn Arg Gln Pro Thr Glu Gly Asn 820 825
830 Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys
Asp Ala Ser 835 840 845
Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850
855 860 Met Ala Gln Lys
Phe Arg Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg 865 870
875 880 Lys Asp Lys Asp Val His Ile Ile Leu
Asp Lys Leu Ser Asn Val Thr 885 890
895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp
Ile Lys 900 905 910
Lys Val Asn Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr
915 920 925 Leu Ile Val Ser
Ala Val Thr Pro Asp Leu Asn Met Thr Arg Gln Lys 930
935 940 Ala Ala Thr Pro Val Thr Ile Asn
Val Thr Ile Asn Gly Lys Trp Gln 945 950
955 960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val
Ser Gly Asp Asn 965 970
975 Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys
980 985 990 Leu Ser Pro
Leu Pro Cys Pro Gly Ala Cys Val Cys Tyr Asn Glu Pro 995
1000 1005 Lys Val Thr Thr Ser Cys
Pro Gln Gln Gly Leu Gln Ala Val Pro 1010 1015
1020 Val Gly Ile Pro Ala Ala Ser Gln Arg Ile Phe
Leu His Gly Asn 1025 1030 1035
Arg Ile Ser His Val Pro Ala Ala Ser Phe Arg Ala Cys Arg Asn
1040 1045 1050 Leu Thr Ile
Leu Trp Leu His Ser Asn Val Leu Ala Arg Ile Asp 1055
1060 1065 Ala Ala Ala Phe Thr Gly Leu Ala
Leu Leu Glu Gln Leu Asp Leu 1070 1075
1080 Ser Asp Asn Ala Gln Leu Arg Ser Val Asp Pro Ala Thr
Phe His 1085 1090 1095
Gly Leu Gly Arg Leu His Thr Leu His Leu Asp Arg Cys Gly Leu 1100
1105 1110 Gln Glu Leu Gly Pro
Gly Leu Phe Arg Gly Leu Ala Ala Leu Gln 1115 1120
1125 Tyr Leu Tyr Leu Gln Asp Asn Ala Leu Gln
Ala Leu Pro Asp Asp 1130 1135 1140
Thr Phe Arg Asp Leu Gly Asn Leu Thr His Leu Phe Leu His Gly
1145 1150 1155 Asn Arg
Ile Ser Ser Val Pro Glu Arg Ala Phe Arg Gly Leu His 1160
1165 1170 Ser Leu Asp Arg Leu Leu Leu
His Gln Asn Arg Val Ala His Val 1175 1180
1185 His Pro His Ala Phe Arg Asp Leu Gly Arg Leu Met
Thr Leu Tyr 1190 1195 1200
Leu Phe Ala Asn Asn Leu Ser Ala Leu Pro Thr Glu Ala Leu Ala 1205
1210 1215 Pro Leu Arg Ala Leu
Gln Tyr Leu Arg Leu Asn Asp Asn Pro Trp 1220 1225
1230 Val Cys Asp Cys Arg Ala Arg Pro Leu Trp
Ala Trp Leu Gln Lys 1235 1240 1245
Phe Arg Gly Ser Ser Ser Glu Val Pro Cys Ser Leu Pro Gln Arg
1250 1255 1260 Leu Ala
Gly Arg Asp Leu Lys Arg Leu Ala Ala Asn Asp Leu Gln 1265
1270 1275 Gly Cys Ala Val 1280
301287PRTArtificialSynthetic polypeptide, C terminal fusion
chimera between NgR(sub)27-311 and chondroitinase ABCI protein with
a peptide spacer or linking group 30Ala Thr Ser Asn Pro Ala Phe Asp Pro
Lys Asn Leu Met Gln Ser Glu 1 5 10
15 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser
Ser Asp 20 25 30
Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn
35 40 45 Gln Ser Leu Leu
Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50
55 60 Lys Lys Leu Ile Val Pro Thr Asp
Lys Glu Ala Ser Lys Ala Trp Gly 65 70
75 80 Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr
Asn Glu Lys Pro 85 90
95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr
100 105 110 Ser Glu Ala
Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp 115
120 125 Arg Thr Val Gly Val Ser Leu Asn
Asn Asp Leu Glu Asn Arg Glu Met 130 135
140 Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln
Asp Ser Ile 145 150 155
160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro
165 170 175 Ser Asn Val Ser
Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser 180
185 190 Val Asp Asp Ala Arg Tyr Gln Trp Ser
Asp Tyr Gln Val Lys Thr Arg 195 200
205 Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln
Leu Pro 210 215 220
Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu 225
230 235 240 Ile Asn Glu Phe Val
Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu 245
250 255 Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe
Asp Ala Leu Asn Thr His 260 265
270 Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp
Lys 275 280 285 Gln
Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln 290
295 300 Leu Phe Asp Asn Tyr Val
Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305 310
315 320 Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp
Pro Thr Gln Lys Ala 325 330
335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln
340 345 350 Gly Phe
Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr 355
360 365 Ser Ser Arg Trp Trp Tyr Ile
Ser Thr Leu Leu Met Ser Asp Ala Leu 370 375
380 Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser
Leu Leu Trp Tyr 385 390 395
400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser
405 410 415 Ser Asp Leu
Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu 420
425 430 Leu Leu Leu Glu Pro Asp Asp Gln
Lys Arg Ile Asn Leu Val Asn Thr 435 440
445 Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro
Pro Gly Gly 450 455 460
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn 465
470 475 480 Tyr Pro Gly Tyr
Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile 485
490 495 Tyr Leu Leu Arg Asp Thr Pro Phe Ser
Val Gly Glu Ser Gly Trp Asn 500 505
510 Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn
Pro Glu 515 520 525
Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu 530
535 540 Lys Ser Val Ala Gln
Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545 550
555 560 Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr
Leu Ala Ile Ser Asp Lys 565 570
575 Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro
Ala 580 585 590 Ser
Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile 595
600 605 His Arg Trp Gln Asp Lys
Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610 615
620 Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn
Arg Tyr Gly Arg Tyr 625 630 635
640 Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser
645 650 655 Gln Gly
Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala 660
665 670 Thr Thr Ile His Leu Pro Leu
Lys Asp Leu Asp Ser Pro Lys Pro His 675 680
685 Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly
Thr Ser Ser Leu 690 695 700
Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn 705
710 715 720 Leu Glu Arg
Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala 725
730 735 Ala Asp Asn His Leu Ile Phe Ile
Gly Ser Asn Ile Asn Ser Ser Asp 740 745
750 Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His Ala
Ile Thr Pro 755 760 765
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro 770
775 780 Tyr Gln Thr Thr
Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785 790
795 800 Asn Gly Tyr Leu Ile Thr Gln Ala Glu
Lys Val Asn Val Ser Arg Gln 805 810
815 His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu
Gly Asn 820 825 830
Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser
835 840 845 Tyr Glu Tyr Met
Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850
855 860 Met Ala Gln Lys Phe Arg Glu Asn
Asn Gly Leu Tyr Gln Val Leu Arg 865 870
875 880 Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu
Ser Asn Val Thr 885 890
895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys
900 905 910 Lys Val Asn
Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr 915
920 925 Leu Ile Val Ser Ala Val Thr Pro
Asp Leu Asn Met Thr Arg Gln Lys 930 935
940 Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn Gly
Lys Trp Gln 945 950 955
960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn
965 970 975 Thr Glu Leu Thr
Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys 980
985 990 Leu Ser Pro Leu Pro Gly Gly Gly
Gly Gly Cys Pro Gly Ala Cys Val 995 1000
1005 Cys Tyr Asn Glu Pro Lys Val Thr Thr Ser Cys
Pro Gln Gln Gly 1010 1015 1020
Leu Gln Ala Val Pro Val Gly Ile Pro Ala Ala Ser Gln Arg Ile
1025 1030 1035 Phe Leu His
Gly Asn Arg Ile Ser His Val Pro Ala Ala Ser Phe 1040
1045 1050 Arg Ala Cys Arg Asn Leu Thr Ile
Leu Trp Leu His Ser Asn Val 1055 1060
1065 Leu Ala Arg Ile Asp Ala Ala Ala Phe Thr Gly Leu Ala
Leu Leu 1070 1075 1080
Glu Gln Leu Asp Leu Ser Asp Asn Ala Gln Leu Arg Ser Val Asp 1085
1090 1095 Pro Ala Thr Phe His
Gly Leu Gly Arg Leu His Thr Leu His Leu 1100 1105
1110 Asp Arg Cys Gly Leu Gln Glu Leu Gly Pro
Gly Leu Phe Arg Gly 1115 1120 1125
Leu Ala Ala Leu Gln Tyr Leu Tyr Leu Gln Asp Asn Ala Leu Gln
1130 1135 1140 Ala Leu
Pro Asp Asp Thr Phe Arg Asp Leu Gly Asn Leu Thr His 1145
1150 1155 Leu Phe Leu His Gly Asn Arg
Ile Ser Ser Val Pro Glu Arg Ala 1160 1165
1170 Phe Arg Gly Leu His Ser Leu Asp Arg Leu Leu Leu
His Gln Asn 1175 1180 1185
Arg Val Ala His Val His Pro His Ala Phe Arg Asp Leu Gly Arg 1190
1195 1200 Leu Met Thr Leu Tyr
Leu Phe Ala Asn Asn Leu Ser Ala Leu Pro 1205 1210
1215 Thr Glu Ala Leu Ala Pro Leu Arg Ala Leu
Gln Tyr Leu Arg Leu 1220 1225 1230
Asn Asp Asn Pro Trp Val Cys Asp Cys Arg Ala Arg Pro Leu Trp
1235 1240 1245 Ala Trp
Leu Gln Lys Phe Arg Gly Ser Ser Ser Glu Val Pro Cys 1250
1255 1260 Ser Leu Pro Gln Arg Leu Ala
Gly Arg Asp Leu Lys Arg Leu Ala 1265 1270
1275 Ala Asn Asp Leu Gln Gly Cys Ala Val 1280
1285 312254PRTArtificialSynthetic polypeptide, N
terminal chimeric fusion between extracellular domain L1 and
chondroitinase ABCI protein without a spacer or linking peptide
31Met Val Val Ala Leu Arg Tyr Val Trp Pro Leu Leu Leu Cys Ser Pro 1
5 10 15 Cys Leu Leu Ile
Gln Ile Pro Glu Glu Tyr Glu Gly His His Val Met 20
25 30 Glu Pro Pro Val Ile Thr Glu Gln Ser
Pro Arg Arg Leu Val Val Phe 35 40
45 Pro Thr Asp Asp Ile Ser Leu Lys Cys Glu Ala Ser Gly Lys
Pro Glu 50 55 60
Val Gln Phe Arg Trp Thr Arg Asp Gly Val His Phe Lys Pro Lys Glu 65
70 75 80 Glu Leu Gly Val Thr
Val Tyr Gln Ser Pro His Ser Gly Ser Phe Thr 85
90 95 Ile Thr Gly Asn Asn Ser Asn Phe Ala Gln
Arg Phe Gln Gly Ile Tyr 100 105
110 Arg Cys Phe Ala Ser Asn Lys Leu Gly Thr Ala Met Ser His Glu
Ile 115 120 125 Arg
Leu Met Ala Glu Gly Ala Pro Lys Trp Pro Lys Glu Thr Val Lys 130
135 140 Pro Val Glu Val Glu Glu
Gly Glu Ser Val Val Leu Pro Cys Asn Pro 145 150
155 160 Pro Pro Ser Ala Glu Pro Leu Arg Ile Tyr Trp
Met Asn Ser Lys Ile 165 170
175 Leu His Ile Lys Gln Asp Glu Arg Val Thr Met Gly Gln Asn Gly Asn
180 185 190 Leu Tyr
Phe Ala Asn Val Leu Thr Ser Asp Asn His Ser Asp Tyr Ile 195
200 205 Cys His Ala His Phe Pro Gly
Thr Arg Thr Ile Ile Gln Lys Glu Pro 210 215
220 Ile Asp Leu Arg Val Lys Ala Thr Asn Ser Met Ile
Asp Arg Lys Pro 225 230 235
240 Arg Leu Leu Phe Pro Thr Asn Ser Ser Ser His Leu Val Ala Leu Gln
245 250 255 Gly Gln Pro
Leu Val Leu Glu Cys Ile Ala Glu Gly Phe Pro Thr Pro 260
265 270 Thr Ile Lys Trp Leu Arg Pro Ser
Gly Pro Met Pro Ala Asp Arg Val 275 280
285 Thr Tyr Gln Asn His Asn Lys Thr Leu Gln Leu Leu Lys
Val Gly Glu 290 295 300
Glu Asp Asp Gly Glu Tyr Arg Cys Leu Ala Glu Asn Ser Leu Gly Ser 305
310 315 320 Ala Arg His Ala
Tyr Tyr Val Thr Val Glu Ala Ala Pro Tyr Trp Leu 325
330 335 His Lys Pro Gln Ser His Leu Tyr Gly
Pro Gly Glu Thr Ala Arg Leu 340 345
350 Asp Cys Gln Val Gln Gly Arg Pro Gln Pro Glu Val Thr Trp
Arg Ile 355 360 365
Asn Gly Ile Pro Val Glu Glu Leu Ala Lys Asp Gln Lys Tyr Arg Ile 370
375 380 Gln Arg Gly Ala Leu
Ile Leu Ser Asn Val Gln Pro Ser Asp Thr Met 385 390
395 400 Val Thr Gln Cys Glu Ala Arg Asn Arg His
Gly Leu Leu Leu Ala Asn 405 410
415 Ala Tyr Ile Tyr Val Val Gln Leu Pro Ala Lys Ile Leu Thr Ala
Asp 420 425 430 Asn
Gln Thr Tyr Met Ala Val Gln Gly Ser Thr Ala Tyr Leu Leu Cys 435
440 445 Lys Ala Phe Gly Ala Pro
Val Pro Ser Val Gln Trp Leu Asp Glu Asp 450 455
460 Gly Thr Thr Val Leu Gln Asp Glu Arg Phe Phe
Pro Tyr Ala Asn Gly 465 470 475
480 Thr Leu Gly Ile Arg Asp Leu Gln Ala Asn Asp Thr Gly Arg Tyr Phe
485 490 495 Cys Leu
Ala Ala Asn Asp Gln Asn Asn Val Thr Ile Met Ala Asn Leu 500
505 510 Lys Val Lys Asp Ala Thr Gln
Ile Thr Gln Gly Pro Arg Ser Thr Ile 515 520
525 Glu Lys Lys Gly Ser Arg Val Thr Phe Thr Cys Gln
Ala Ser Phe Asp 530 535 540
Pro Ser Leu Gln Pro Ser Ile Thr Trp Arg Gly Asp Gly Arg Asp Leu 545
550 555 560 Gln Glu Leu
Gly Asp Ser Asp Lys Tyr Phe Ile Glu Asp Gly Arg Leu 565
570 575 Val Ile His Ser Leu Asp Tyr Ser
Asp Gln Gly Asn Tyr Ser Cys Val 580 585
590 Ala Ser Thr Glu Leu Asp Val Val Glu Ser Arg Ala Gln
Leu Leu Val 595 600 605
Val Gly Ser Pro Gly Pro Val Pro Arg Leu Val Leu Ser Asp Leu His 610
615 620 Leu Leu Thr Gln
Ser Gln Val Arg Val Ser Trp Ser Pro Ala Glu Asp 625 630
635 640 His Asn Ala Pro Ile Glu Lys Tyr Asp
Ile Glu Phe Glu Asp Lys Glu 645 650
655 Met Ala Pro Glu Lys Trp Tyr Ser Leu Gly Lys Val Pro Gly
Asn Gln 660 665 670
Thr Ser Thr Thr Leu Lys Leu Ser Pro Tyr Val His Tyr Thr Phe Arg
675 680 685 Val Thr Ala Ile
Asn Lys Tyr Gly Pro Gly Glu Pro Ser Pro Val Ser 690
695 700 Glu Thr Val Val Thr Pro Glu Ala
Ala Pro Glu Lys Asn Pro Val Asp 705 710
715 720 Val Lys Gly Glu Gly Asn Glu Thr Thr Asn Met Val
Ile Thr Trp Lys 725 730
735 Pro Leu Arg Trp Met Asp Trp Asn Ala Pro Gln Val Gln Tyr Arg Val
740 745 750 Gln Trp Arg
Pro Gln Gly Thr Arg Gly Pro Trp Gln Glu Gln Ile Val 755
760 765 Ser Asp Pro Phe Leu Val Val Ser
Asn Thr Ser Thr Phe Val Pro Tyr 770 775
780 Glu Ile Lys Val Gln Ala Val Asn Ser Gln Gly Lys Gly
Pro Glu Pro 785 790 795
800 Gln Val Thr Ile Gly Tyr Ser Gly Glu Asp Tyr Pro Gln Ala Ile Pro
805 810 815 Glu Leu Glu Gly
Ile Glu Ile Leu Asn Ser Ser Ala Val Leu Val Lys 820
825 830 Trp Arg Pro Val Asp Leu Ala Gln Val
Lys Gly His Leu Arg Gly Tyr 835 840
845 Asn Val Thr Tyr Trp Arg Glu Gly Ser Gln Arg Lys His Ser
Lys Arg 850 855 860
His Ile His Lys Asp His Val Val Val Pro Ala Asn Thr Thr Ser Val 865
870 875 880 Ile Leu Ser Gly Leu
Arg Pro Tyr Ser Ser Tyr His Leu Glu Val Gln 885
890 895 Ala Phe Asn Gly Arg Gly Ser Gly Pro Ala
Ser Glu Phe Thr Phe Ser 900 905
910 Thr Pro Glu Gly Val Pro Gly His Pro Glu Ala Leu His Leu Glu
Cys 915 920 925 Gln
Ser Asn Thr Ser Leu Leu Leu Arg Trp Gln Pro Pro Leu Ser His 930
935 940 Asn Gly Val Leu Thr Gly
Tyr Val Leu Ser Tyr His Pro Leu Asp Glu 945 950
955 960 Gly Gly Lys Gly Gln Leu Ser Phe Asn Leu Arg
Asp Pro Glu Leu Arg 965 970
975 Thr His Asn Leu Thr Asp Leu Ser Pro His Leu Arg Tyr Arg Phe Gln
980 985 990 Leu Gln
Ala Thr Thr Lys Glu Gly Pro Gly Glu Ala Ile Val Arg Glu 995
1000 1005 Gly Gly Thr Met Ala
Leu Ser Gly Ile Ser Asp Phe Gly Asn Ile 1010 1015
1020 Ser Ala Thr Ala Gly Glu Asn Tyr Ser Val
Val Ser Trp Val Pro 1025 1030 1035
Lys Glu Gly Gln Cys Asn Phe Arg Phe His Ile Leu Phe Lys Ala
1040 1045 1050 Leu Gly
Glu Glu Lys Gly Gly Ala Ser Leu Ser Pro Gln Tyr Val 1055
1060 1065 Ser Tyr Asn Gln Ser Ser Tyr
Thr Gln Trp Asp Leu Gln Pro Asp 1070 1075
1080 Thr Asp Tyr Glu Ile His Leu Phe Lys Glu Arg Met
Phe Arg His 1085 1090 1095
Gln Met Ala Val Lys Thr Asn Gly Thr Gly Arg Val Arg Leu Pro 1100
1105 1110 Pro Ala Gly Phe Ala
Thr Glu Gly Trp Phe Ile Gly Phe Val Ser 1115 1120
1125 Ala Ile Ile Leu Leu Leu Leu Val Leu Leu
Ile Leu Cys Phe Ile 1130 1135 1140
Lys Arg Ser Lys Gly Gly Lys Tyr Ser Val Lys Asp Lys Glu Asp
1145 1150 1155 Thr Gln
Val Asp Ser Glu Ala Arg Pro Met Lys Asp Glu Thr Phe 1160
1165 1170 Gly Glu Tyr Arg Ser Leu Glu
Ser Asp Asn Glu Glu Lys Ala Phe 1175 1180
1185 Gly Ser Ser Gln Pro Ser Leu Asn Gly Asp Ile Lys
Pro Leu Gly 1190 1195 1200
Ser Asp Asp Ser Leu Ala Asp Tyr Gly Gly Ser Val Asp Val Gln 1205
1210 1215 Phe Asn Glu Asp Gly
Ser Phe Ile Gly Gln Tyr Ser Gly Lys Lys 1220 1225
1230 Glu Lys Glu Ala Ala Gly Gly Asn Asp Ser
Ser Gly Ala Thr Ser 1235 1240 1245
Pro Ile Asn Pro Ala Val Ala Leu Glu Ala Thr Ser Asn Pro Ala
1250 1255 1260 Phe Asp
Pro Lys Asn Leu Met Gln Ser Glu Ile Tyr His Phe Ala 1265
1270 1275 Gln Asn Asn Pro Leu Ala Asp
Phe Ser Ser Asp Lys Asn Ser Ile 1280 1285
1290 Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn
Gln Ser Leu 1295 1300 1305
Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His Lys Lys 1310
1315 1320 Leu Ile Val Pro Thr
Asp Lys Glu Ala Ser Lys Ala Trp Gly Arg 1325 1330
1335 Ser Ser Thr Pro Val Phe Ser Phe Trp Leu
Tyr Asn Glu Lys Pro 1340 1345 1350
Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser
1355 1360 1365 Thr Ser
Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr 1370
1375 1380 Gly Trp Arg Thr Val Gly Val
Ser Leu Asn Asn Asp Leu Glu Asn 1385 1390
1395 Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser Ser
Asp Gly Thr 1400 1405 1410
Gln Asp Ser Ile Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile 1415
1420 1425 Arg Phe Lys Ala Pro
Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile 1430 1435
1440 Asp Arg Ile Met Phe Ser Val Asp Asp Ala
Arg Tyr Gln Trp Ser 1445 1450 1455
Asp Tyr Gln Val Lys Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe
1460 1465 1470 His Asn
Val Lys Pro Gln Leu Pro Val Thr Pro Glu Asn Leu Ala 1475
1480 1485 Ala Ile Asp Leu Ile Arg Gln
Arg Leu Ile Asn Glu Phe Val Gly 1490 1495
1500 Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu Glu Asn
Ile Ser Lys 1505 1510 1515
Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr His Thr Leu Ala Asn 1520
1525 1530 Gly Gly Thr Gln Gly
Arg His Leu Ile Thr Asp Lys Gln Ile Ile 1535 1540
1545 Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln
Asp Lys Gln Leu Phe 1550 1555 1560
Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe Asn
1565 1570 1575 Ile Ser
Arg Ala Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys Ala 1580
1585 1590 Gln Leu Lys Gln Met Tyr Leu
Leu Met Thr Lys His Leu Leu Asp 1595 1600
1605 Gln Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr
His His Trp 1610 1615 1620
Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser 1625
1630 1635 Asp Ala Leu Lys Glu
Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser 1640 1645
1650 Leu Leu Trp Tyr Ser Arg Glu Phe Lys Ser
Ser Phe Asp Met Lys 1655 1660 1665
Val Ser Ala Asp Ser Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser
1670 1675 1680 Arg Gln
His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln Lys 1685
1690 1695 Arg Ile Asn Leu Val Asn Thr
Phe Ser His Tyr Ile Thr Gly Ala 1700 1705
1710 Leu Thr Gln Val Pro Pro Gly Gly Lys Asp Gly Leu
Arg Pro Asp 1715 1720 1725
Gly Thr Ala Trp Arg His Glu Gly Asn Tyr Pro Gly Tyr Ser Phe 1730
1735 1740 Pro Ala Phe Lys Asn
Ala Ser Gln Leu Ile Tyr Leu Leu Arg Asp 1745 1750
1755 Thr Pro Phe Ser Val Gly Glu Ser Gly Trp
Asn Asn Leu Lys Lys 1760 1765 1770
Ala Met Val Ser Ala Trp Ile Tyr Ser Asn Pro Glu Val Gly Leu
1775 1780 1785 Pro Leu
Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu Lys Ser 1790
1795 1800 Val Ala Gln Gly Tyr Tyr Trp
Leu Ala Met Ser Ala Lys Ser Ser 1805 1810
1815 Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu Ala Ile
Ser Asp Lys 1820 1825 1830
Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro 1835
1840 1845 Ala Ser Leu Pro Gln
Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe 1850 1855
1860 Gly Ile His Arg Trp Gln Asp Lys Met Val
Thr Leu Lys Ala Tyr 1865 1870 1875
Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg
1880 1885 1890 Tyr Gly
Arg Tyr Gln Ser His Gly Val Ala Gln Ile Val Ser Asn 1895
1900 1905 Gly Ser Gln Leu Ser Gln Gly
Tyr Gln Gln Glu Gly Trp Asp Trp 1910 1915
1920 Asn Arg Met Glu Gly Ala Thr Thr Ile His Leu Pro
Leu Lys Asp 1925 1930 1935
Leu Asp Ser Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg 1940
1945 1950 Gly Phe Ser Gly Thr
Ser Ser Leu Glu Gly Gln Tyr Gly Met Met 1955 1960
1965 Ala Phe Asn Leu Ile Tyr Pro Ala Asn Leu
Glu Arg Phe Asp Pro 1970 1975 1980
Asn Phe Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His Leu
1985 1990 1995 Ile Phe
Ile Gly Ser Asn Ile Asn Ser Ser Asp Lys Asn Lys Asn 2000
2005 2010 Val Glu Thr Thr Leu Phe Gln
His Ala Ile Thr Pro Thr Leu Asn 2015 2020
2025 Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met
Pro Tyr Gln 2030 2035 2040
Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly Asn 2045
2050 2055 Gly Tyr Leu Ile Thr
Gln Ala Glu Lys Val Asn Val Ser Arg Gln 2060 2065
2070 His Gln Val Ser Ala Glu Asn Lys Asn Arg
Gln Pro Thr Glu Gly 2075 2080 2085
Asn Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp
2090 2095 2100 Ala Ser
Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys 2105
2110 2115 Met Gly Glu Met Ala Gln Lys
Phe Arg Glu Asn Asn Gly Leu Tyr 2120 2125
2130 Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile
Leu Asp Lys 2135 2140 2145
Leu Ser Asn Val Thr Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile 2150
2155 2160 Glu Asp Lys Trp Ile
Lys Lys Val Asn Lys Pro Ala Ile Val Met 2165 2170
2175 Thr His Arg Gln Lys Asp Thr Leu Ile Val
Ser Ala Val Thr Pro 2180 2185 2190
Asp Leu Asn Met Thr Arg Gln Lys Ala Ala Thr Pro Val Thr Ile
2195 2200 2205 Asn Val
Thr Ile Asn Gly Lys Trp Gln Ser Ala Asp Lys Asn Ser 2210
2215 2220 Glu Val Lys Tyr Gln Val Ser
Gly Asp Asn Thr Glu Leu Thr Phe 2225 2230
2235 Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys Leu
Ser Pro Leu 2240 2245 2250
Pro 322259PRTArtificialSynthetic polypeptide, N terminal fusion
chimera between extracellular domain L1 and chondroitinase ABCI
protein with a spacer or linking peptide 32Met Val Val Ala Leu Arg Tyr
Val Trp Pro Leu Leu Leu Cys Ser Pro 1 5
10 15 Cys Leu Leu Ile Gln Ile Pro Glu Glu Tyr Glu
Gly His His Val Met 20 25
30 Glu Pro Pro Val Ile Thr Glu Gln Ser Pro Arg Arg Leu Val Val
Phe 35 40 45 Pro
Thr Asp Asp Ile Ser Leu Lys Cys Glu Ala Ser Gly Lys Pro Glu 50
55 60 Val Gln Phe Arg Trp Thr
Arg Asp Gly Val His Phe Lys Pro Lys Glu 65 70
75 80 Glu Leu Gly Val Thr Val Tyr Gln Ser Pro His
Ser Gly Ser Phe Thr 85 90
95 Ile Thr Gly Asn Asn Ser Asn Phe Ala Gln Arg Phe Gln Gly Ile Tyr
100 105 110 Arg Cys
Phe Ala Ser Asn Lys Leu Gly Thr Ala Met Ser His Glu Ile 115
120 125 Arg Leu Met Ala Glu Gly Ala
Pro Lys Trp Pro Lys Glu Thr Val Lys 130 135
140 Pro Val Glu Val Glu Glu Gly Glu Ser Val Val Leu
Pro Cys Asn Pro 145 150 155
160 Pro Pro Ser Ala Glu Pro Leu Arg Ile Tyr Trp Met Asn Ser Lys Ile
165 170 175 Leu His Ile
Lys Gln Asp Glu Arg Val Thr Met Gly Gln Asn Gly Asn 180
185 190 Leu Tyr Phe Ala Asn Val Leu Thr
Ser Asp Asn His Ser Asp Tyr Ile 195 200
205 Cys His Ala His Phe Pro Gly Thr Arg Thr Ile Ile Gln
Lys Glu Pro 210 215 220
Ile Asp Leu Arg Val Lys Ala Thr Asn Ser Met Ile Asp Arg Lys Pro 225
230 235 240 Arg Leu Leu Phe
Pro Thr Asn Ser Ser Ser His Leu Val Ala Leu Gln 245
250 255 Gly Gln Pro Leu Val Leu Glu Cys Ile
Ala Glu Gly Phe Pro Thr Pro 260 265
270 Thr Ile Lys Trp Leu Arg Pro Ser Gly Pro Met Pro Ala Asp
Arg Val 275 280 285
Thr Tyr Gln Asn His Asn Lys Thr Leu Gln Leu Leu Lys Val Gly Glu 290
295 300 Glu Asp Asp Gly Glu
Tyr Arg Cys Leu Ala Glu Asn Ser Leu Gly Ser 305 310
315 320 Ala Arg His Ala Tyr Tyr Val Thr Val Glu
Ala Ala Pro Tyr Trp Leu 325 330
335 His Lys Pro Gln Ser His Leu Tyr Gly Pro Gly Glu Thr Ala Arg
Leu 340 345 350 Asp
Cys Gln Val Gln Gly Arg Pro Gln Pro Glu Val Thr Trp Arg Ile 355
360 365 Asn Gly Ile Pro Val Glu
Glu Leu Ala Lys Asp Gln Lys Tyr Arg Ile 370 375
380 Gln Arg Gly Ala Leu Ile Leu Ser Asn Val Gln
Pro Ser Asp Thr Met 385 390 395
400 Val Thr Gln Cys Glu Ala Arg Asn Arg His Gly Leu Leu Leu Ala Asn
405 410 415 Ala Tyr
Ile Tyr Val Val Gln Leu Pro Ala Lys Ile Leu Thr Ala Asp 420
425 430 Asn Gln Thr Tyr Met Ala Val
Gln Gly Ser Thr Ala Tyr Leu Leu Cys 435 440
445 Lys Ala Phe Gly Ala Pro Val Pro Ser Val Gln Trp
Leu Asp Glu Asp 450 455 460
Gly Thr Thr Val Leu Gln Asp Glu Arg Phe Phe Pro Tyr Ala Asn Gly 465
470 475 480 Thr Leu Gly
Ile Arg Asp Leu Gln Ala Asn Asp Thr Gly Arg Tyr Phe 485
490 495 Cys Leu Ala Ala Asn Asp Gln Asn
Asn Val Thr Ile Met Ala Asn Leu 500 505
510 Lys Val Lys Asp Ala Thr Gln Ile Thr Gln Gly Pro Arg
Ser Thr Ile 515 520 525
Glu Lys Lys Gly Ser Arg Val Thr Phe Thr Cys Gln Ala Ser Phe Asp 530
535 540 Pro Ser Leu Gln
Pro Ser Ile Thr Trp Arg Gly Asp Gly Arg Asp Leu 545 550
555 560 Gln Glu Leu Gly Asp Ser Asp Lys Tyr
Phe Ile Glu Asp Gly Arg Leu 565 570
575 Val Ile His Ser Leu Asp Tyr Ser Asp Gln Gly Asn Tyr Ser
Cys Val 580 585 590
Ala Ser Thr Glu Leu Asp Val Val Glu Ser Arg Ala Gln Leu Leu Val
595 600 605 Val Gly Ser Pro
Gly Pro Val Pro Arg Leu Val Leu Ser Asp Leu His 610
615 620 Leu Leu Thr Gln Ser Gln Val Arg
Val Ser Trp Ser Pro Ala Glu Asp 625 630
635 640 His Asn Ala Pro Ile Glu Lys Tyr Asp Ile Glu Phe
Glu Asp Lys Glu 645 650
655 Met Ala Pro Glu Lys Trp Tyr Ser Leu Gly Lys Val Pro Gly Asn Gln
660 665 670 Thr Ser Thr
Thr Leu Lys Leu Ser Pro Tyr Val His Tyr Thr Phe Arg 675
680 685 Val Thr Ala Ile Asn Lys Tyr Gly
Pro Gly Glu Pro Ser Pro Val Ser 690 695
700 Glu Thr Val Val Thr Pro Glu Ala Ala Pro Glu Lys Asn
Pro Val Asp 705 710 715
720 Val Lys Gly Glu Gly Asn Glu Thr Thr Asn Met Val Ile Thr Trp Lys
725 730 735 Pro Leu Arg Trp
Met Asp Trp Asn Ala Pro Gln Val Gln Tyr Arg Val 740
745 750 Gln Trp Arg Pro Gln Gly Thr Arg Gly
Pro Trp Gln Glu Gln Ile Val 755 760
765 Ser Asp Pro Phe Leu Val Val Ser Asn Thr Ser Thr Phe Val
Pro Tyr 770 775 780
Glu Ile Lys Val Gln Ala Val Asn Ser Gln Gly Lys Gly Pro Glu Pro 785
790 795 800 Gln Val Thr Ile Gly
Tyr Ser Gly Glu Asp Tyr Pro Gln Ala Ile Pro 805
810 815 Glu Leu Glu Gly Ile Glu Ile Leu Asn Ser
Ser Ala Val Leu Val Lys 820 825
830 Trp Arg Pro Val Asp Leu Ala Gln Val Lys Gly His Leu Arg Gly
Tyr 835 840 845 Asn
Val Thr Tyr Trp Arg Glu Gly Ser Gln Arg Lys His Ser Lys Arg 850
855 860 His Ile His Lys Asp His
Val Val Val Pro Ala Asn Thr Thr Ser Val 865 870
875 880 Ile Leu Ser Gly Leu Arg Pro Tyr Ser Ser Tyr
His Leu Glu Val Gln 885 890
895 Ala Phe Asn Gly Arg Gly Ser Gly Pro Ala Ser Glu Phe Thr Phe Ser
900 905 910 Thr Pro
Glu Gly Val Pro Gly His Pro Glu Ala Leu His Leu Glu Cys 915
920 925 Gln Ser Asn Thr Ser Leu Leu
Leu Arg Trp Gln Pro Pro Leu Ser His 930 935
940 Asn Gly Val Leu Thr Gly Tyr Val Leu Ser Tyr His
Pro Leu Asp Glu 945 950 955
960 Gly Gly Lys Gly Gln Leu Ser Phe Asn Leu Arg Asp Pro Glu Leu Arg
965 970 975 Thr His Asn
Leu Thr Asp Leu Ser Pro His Leu Arg Tyr Arg Phe Gln 980
985 990 Leu Gln Ala Thr Thr Lys Glu Gly
Pro Gly Glu Ala Ile Val Arg Glu 995 1000
1005 Gly Gly Thr Met Ala Leu Ser Gly Ile Ser Asp
Phe Gly Asn Ile 1010 1015 1020
Ser Ala Thr Ala Gly Glu Asn Tyr Ser Val Val Ser Trp Val Pro
1025 1030 1035 Lys Glu Gly
Gln Cys Asn Phe Arg Phe His Ile Leu Phe Lys Ala 1040
1045 1050 Leu Gly Glu Glu Lys Gly Gly Ala
Ser Leu Ser Pro Gln Tyr Val 1055 1060
1065 Ser Tyr Asn Gln Ser Ser Tyr Thr Gln Trp Asp Leu Gln
Pro Asp 1070 1075 1080
Thr Asp Tyr Glu Ile His Leu Phe Lys Glu Arg Met Phe Arg His 1085
1090 1095 Gln Met Ala Val Lys
Thr Asn Gly Thr Gly Arg Val Arg Leu Pro 1100 1105
1110 Pro Ala Gly Phe Ala Thr Glu Gly Trp Phe
Ile Gly Phe Val Ser 1115 1120 1125
Ala Ile Ile Leu Leu Leu Leu Val Leu Leu Ile Leu Cys Phe Ile
1130 1135 1140 Lys Arg
Ser Lys Gly Gly Lys Tyr Ser Val Lys Asp Lys Glu Asp 1145
1150 1155 Thr Gln Val Asp Ser Glu Ala
Arg Pro Met Lys Asp Glu Thr Phe 1160 1165
1170 Gly Glu Tyr Arg Ser Leu Glu Ser Asp Asn Glu Glu
Lys Ala Phe 1175 1180 1185
Gly Ser Ser Gln Pro Ser Leu Asn Gly Asp Ile Lys Pro Leu Gly 1190
1195 1200 Ser Asp Asp Ser Leu
Ala Asp Tyr Gly Gly Ser Val Asp Val Gln 1205 1210
1215 Phe Asn Glu Asp Gly Ser Phe Ile Gly Gln
Tyr Ser Gly Lys Lys 1220 1225 1230
Glu Lys Glu Ala Ala Gly Gly Asn Asp Ser Ser Gly Ala Thr Ser
1235 1240 1245 Pro Ile
Asn Pro Ala Val Ala Leu Glu Gly Gly Gly Gly Gly Ala 1250
1255 1260 Thr Ser Asn Pro Ala Phe Asp
Pro Lys Asn Leu Met Gln Ser Glu 1265 1270
1275 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala Asp
Phe Ser Ser 1280 1285 1290
Asp Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met 1295
1300 1305 Gly Asn Gln Ser Leu
Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe 1310 1315
1320 Thr Leu His Lys Lys Leu Ile Val Pro Thr
Asp Lys Glu Ala Ser 1325 1330 1335
Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu
1340 1345 1350 Tyr Asn
Glu Lys Pro Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly 1355
1360 1365 Glu Lys Leu Ile Ser Thr Ser
Glu Ala Gln Ala Gly Phe Lys Val 1370 1375
1380 Lys Leu Asp Phe Thr Gly Trp Arg Thr Val Gly Val
Ser Leu Asn 1385 1390 1395
Asn Asp Leu Glu Asn Arg Glu Met Thr Leu Asn Ala Thr Asn Thr 1400
1405 1410 Ser Ser Asp Gly Thr
Gln Asp Ser Ile Gly Arg Ser Leu Gly Ala 1415 1420
1425 Lys Val Asp Ser Ile Arg Phe Lys Ala Pro
Ser Asn Val Ser Gln 1430 1435 1440
Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala
1445 1450 1455 Arg Tyr
Gln Trp Ser Asp Tyr Gln Val Lys Thr Arg Leu Ser Glu 1460
1465 1470 Pro Glu Ile Gln Phe His Asn
Val Lys Pro Gln Leu Pro Val Thr 1475 1480
1485 Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln
Arg Leu Ile 1490 1495 1500
Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu 1505
1510 1515 Glu Asn Ile Ser Lys
Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr 1520 1525
1530 His Thr Leu Ala Asn Gly Gly Thr Gln Gly
Arg His Leu Ile Thr 1535 1540 1545
Asp Lys Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln
1550 1555 1560 Asp Lys
Gln Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr 1565
1570 1575 Thr Leu Met Phe Asn Ile Ser
Arg Ala Tyr Val Leu Glu Lys Asp 1580 1585
1590 Pro Thr Gln Lys Ala Gln Leu Lys Gln Met Tyr Leu
Leu Met Thr 1595 1600 1605
Lys His Leu Leu Asp Gln Gly Phe Val Lys Gly Ser Ala Leu Val 1610
1615 1620 Thr Thr His His Trp
Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser 1625 1630
1635 Thr Leu Leu Met Ser Asp Ala Leu Lys Glu
Ala Asn Leu Gln Thr 1640 1645 1650
Gln Val Tyr Asp Ser Leu Leu Trp Tyr Ser Arg Glu Phe Lys Ser
1655 1660 1665 Ser Phe
Asp Met Lys Val Ser Ala Asp Ser Ser Asp Leu Asp Tyr 1670
1675 1680 Phe Asn Thr Leu Ser Arg Gln
His Leu Ala Leu Leu Leu Leu Glu 1685 1690
1695 Pro Asp Asp Gln Lys Arg Ile Asn Leu Val Asn Thr
Phe Ser His 1700 1705 1710
Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro Gly Gly Lys Asp 1715
1720 1725 Gly Leu Arg Pro Asp
Gly Thr Ala Trp Arg His Glu Gly Asn Tyr 1730 1735
1740 Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn
Ala Ser Gln Leu Ile 1745 1750 1755
Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly Trp
1760 1765 1770 Asn Asn
Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn 1775
1780 1785 Pro Glu Val Gly Leu Pro Leu
Ala Gly Arg His Pro Phe Asn Ser 1790 1795
1800 Pro Ser Leu Lys Ser Val Ala Gln Gly Tyr Tyr Trp
Leu Ala Met 1805 1810 1815
Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu 1820
1825 1830 Ala Ile Ser Asp Lys
Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly 1835 1840
1845 Glu Thr Ile Thr Pro Ala Ser Leu Pro Gln
Gly Phe Tyr Ala Phe 1850 1855 1860
Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met Val
1865 1870 1875 Thr Leu
Lys Ala Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr 1880
1885 1890 Asn Lys Asp Asn Arg Tyr Gly
Arg Tyr Gln Ser His Gly Val Ala 1895 1900
1905 Gln Ile Val Ser Asn Gly Ser Gln Leu Ser Gln Gly
Tyr Gln Gln 1910 1915 1920
Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile His 1925
1930 1935 Leu Pro Leu Lys Asp
Leu Asp Ser Pro Lys Pro His Thr Leu Met 1940 1945
1950 Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr
Ser Ser Leu Glu Gly 1955 1960 1965
Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn Leu
1970 1975 1980 Glu Arg
Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala 1985
1990 1995 Ala Asp Asn His Leu Ile Phe
Ile Gly Ser Asn Ile Asn Ser Ser 2000 2005
2010 Asp Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln
His Ala Ile 2015 2020 2025
Thr Pro Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu 2030
2035 2040 Asn Met Pro Tyr Gln
Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile 2045 2050
2055 Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr
Gln Ala Glu Lys Val 2060 2065 2070
Asn Val Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys Asn Arg
2075 2080 2085 Gln Pro
Thr Glu Gly Asn Phe Ser Ser Ala Trp Ile Asp His Ser 2090
2095 2100 Thr Arg Pro Lys Asp Ala Ser
Tyr Glu Tyr Met Val Phe Leu Asp 2105 2110
2115 Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys
Phe Arg Glu 2120 2125 2130
Asn Asn Gly Leu Tyr Gln Val Leu Arg Lys Asp Lys Asp Val His 2135
2140 2145 Ile Ile Leu Asp Lys
Leu Ser Asn Val Thr Gly Tyr Ala Phe Tyr 2150 2155
2160 Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile
Lys Lys Val Asn Lys 2165 2170 2175
Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr Leu Ile Val
2180 2185 2190 Ser Ala
Val Thr Pro Asp Leu Asn Met Thr Arg Gln Lys Ala Ala 2195
2200 2205 Thr Pro Val Thr Ile Asn Val
Thr Ile Asn Gly Lys Trp Gln Ser 2210 2215
2220 Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser
Gly Asp Asn 2225 2230 2235
Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile 2240
2245 2250 Lys Leu Ser Pro Leu
Pro 2255 332254PRTArtificialSynthetic polypeptide, C
terminal fusion chimera between extracellular domain L1 and
chondroitinase ABCI protein without a spacer or linking peptide
33Ala Thr Ser Asn Pro Ala Phe Asp Pro Lys Asn Leu Met Gln Ser Glu 1
5 10 15 Ile Tyr His Phe
Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser Ser Asp 20
25 30 Lys Asn Ser Ile Leu Thr Leu Ser Asp
Lys Arg Ser Ile Met Gly Asn 35 40
45 Gln Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr
Leu His 50 55 60
Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly 65
70 75 80 Arg Ser Ser Thr Pro
Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys Pro 85
90 95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly
Glu Lys Leu Ile Ser Thr 100 105
110 Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly
Trp 115 120 125 Arg
Thr Val Gly Val Ser Leu Asn Asn Asp Leu Glu Asn Arg Glu Met 130
135 140 Thr Leu Asn Ala Thr Asn
Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile 145 150
155 160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile
Arg Phe Lys Ala Pro 165 170
175 Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser
180 185 190 Val Asp
Asp Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val Lys Thr Arg 195
200 205 Leu Ser Glu Pro Glu Ile Gln
Phe His Asn Val Lys Pro Gln Leu Pro 210 215
220 Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile
Arg Gln Arg Leu 225 230 235
240 Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu
245 250 255 Glu Asn Ile
Ser Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr His 260
265 270 Thr Leu Ala Asn Gly Gly Thr Gln
Gly Arg His Leu Ile Thr Asp Lys 275 280
285 Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln
Asp Lys Gln 290 295 300
Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305
310 315 320 Asn Ile Ser Arg
Ala Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys Ala 325
330 335 Gln Leu Lys Gln Met Tyr Leu Leu Met
Thr Lys His Leu Leu Asp Gln 340 345
350 Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp
Gly Tyr 355 360 365
Ser Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser Asp Ala Leu 370
375 380 Lys Glu Ala Asn Leu
Gln Thr Gln Val Tyr Asp Ser Leu Leu Trp Tyr 385 390
395 400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met
Lys Val Ser Ala Asp Ser 405 410
415 Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala
Leu 420 425 430 Leu
Leu Leu Glu Pro Asp Asp Gln Lys Arg Ile Asn Leu Val Asn Thr 435
440 445 Phe Ser His Tyr Ile Thr
Gly Ala Leu Thr Gln Val Pro Pro Gly Gly 450 455
460 Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp
Arg His Glu Gly Asn 465 470 475
480 Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile
485 490 495 Tyr Leu
Leu Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly Trp Asn 500
505 510 Asn Leu Lys Lys Ala Met Val
Ser Ala Trp Ile Tyr Ser Asn Pro Glu 515 520
525 Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn
Ser Pro Ser Leu 530 535 540
Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545
550 555 560 Ser Pro Asp
Lys Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser Asp Lys 565
570 575 Thr Gln Asn Glu Ser Thr Ala Ile
Phe Gly Glu Thr Ile Thr Pro Ala 580 585
590 Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala
Phe Gly Ile 595 600 605
His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610
615 620 Val Trp Ser Ser
Glu Ile Tyr Asn Lys Asp Asn Arg Tyr Gly Arg Tyr 625 630
635 640 Gln Ser His Gly Val Ala Gln Ile Val
Ser Asn Gly Ser Gln Leu Ser 645 650
655 Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu
Gly Ala 660 665 670
Thr Thr Ile His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His
675 680 685 Thr Leu Met Gln
Arg Gly Glu Arg Gly Phe Ser Gly Thr Ser Ser Leu 690
695 700 Glu Gly Gln Tyr Gly Met Met Ala
Phe Asn Leu Ile Tyr Pro Ala Asn 705 710
715 720 Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys
Ser Val Leu Ala 725 730
735 Ala Asp Asn His Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp
740 745 750 Lys Asn Lys
Asn Val Glu Thr Thr Leu Phe Gln His Ala Ile Thr Pro 755
760 765 Thr Leu Asn Thr Leu Trp Ile Asn
Gly Gln Lys Ile Glu Asn Met Pro 770 775
780 Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp
Ser Asn Gly 785 790 795
800 Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln
805 810 815 His Gln Val Ser
Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu Gly Asn 820
825 830 Phe Ser Ser Ala Trp Ile Asp His Ser
Thr Arg Pro Lys Asp Ala Ser 835 840
845 Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys Met
Gly Glu 850 855 860
Met Ala Gln Lys Phe Arg Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg 865
870 875 880 Lys Asp Lys Asp Val
His Ile Ile Leu Asp Lys Leu Ser Asn Val Thr 885
890 895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile
Glu Asp Lys Trp Ile Lys 900 905
910 Lys Val Asn Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp
Thr 915 920 925 Leu
Ile Val Ser Ala Val Thr Pro Asp Leu Asn Met Thr Arg Gln Lys 930
935 940 Ala Ala Thr Pro Val Thr
Ile Asn Val Thr Ile Asn Gly Lys Trp Gln 945 950
955 960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln
Val Ser Gly Asp Asn 965 970
975 Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys
980 985 990 Leu Ser
Pro Leu Pro Met Val Val Ala Leu Arg Tyr Val Trp Pro Leu 995
1000 1005 Leu Leu Cys Ser Pro
Cys Leu Leu Ile Gln Ile Pro Glu Glu Tyr 1010 1015
1020 Glu Gly His His Val Met Glu Pro Pro Val
Ile Thr Glu Gln Ser 1025 1030 1035
Pro Arg Arg Leu Val Val Phe Pro Thr Asp Asp Ile Ser Leu Lys
1040 1045 1050 Cys Glu
Ala Ser Gly Lys Pro Glu Val Gln Phe Arg Trp Thr Arg 1055
1060 1065 Asp Gly Val His Phe Lys Pro
Lys Glu Glu Leu Gly Val Thr Val 1070 1075
1080 Tyr Gln Ser Pro His Ser Gly Ser Phe Thr Ile Thr
Gly Asn Asn 1085 1090 1095
Ser Asn Phe Ala Gln Arg Phe Gln Gly Ile Tyr Arg Cys Phe Ala 1100
1105 1110 Ser Asn Lys Leu Gly
Thr Ala Met Ser His Glu Ile Arg Leu Met 1115 1120
1125 Ala Glu Gly Ala Pro Lys Trp Pro Lys Glu
Thr Val Lys Pro Val 1130 1135 1140
Glu Val Glu Glu Gly Glu Ser Val Val Leu Pro Cys Asn Pro Pro
1145 1150 1155 Pro Ser
Ala Glu Pro Leu Arg Ile Tyr Trp Met Asn Ser Lys Ile 1160
1165 1170 Leu His Ile Lys Gln Asp Glu
Arg Val Thr Met Gly Gln Asn Gly 1175 1180
1185 Asn Leu Tyr Phe Ala Asn Val Leu Thr Ser Asp Asn
His Ser Asp 1190 1195 1200
Tyr Ile Cys His Ala His Phe Pro Gly Thr Arg Thr Ile Ile Gln 1205
1210 1215 Lys Glu Pro Ile Asp
Leu Arg Val Lys Ala Thr Asn Ser Met Ile 1220 1225
1230 Asp Arg Lys Pro Arg Leu Leu Phe Pro Thr
Asn Ser Ser Ser His 1235 1240 1245
Leu Val Ala Leu Gln Gly Gln Pro Leu Val Leu Glu Cys Ile Ala
1250 1255 1260 Glu Gly
Phe Pro Thr Pro Thr Ile Lys Trp Leu Arg Pro Ser Gly 1265
1270 1275 Pro Met Pro Ala Asp Arg Val
Thr Tyr Gln Asn His Asn Lys Thr 1280 1285
1290 Leu Gln Leu Leu Lys Val Gly Glu Glu Asp Asp Gly
Glu Tyr Arg 1295 1300 1305
Cys Leu Ala Glu Asn Ser Leu Gly Ser Ala Arg His Ala Tyr Tyr 1310
1315 1320 Val Thr Val Glu Ala
Ala Pro Tyr Trp Leu His Lys Pro Gln Ser 1325 1330
1335 His Leu Tyr Gly Pro Gly Glu Thr Ala Arg
Leu Asp Cys Gln Val 1340 1345 1350
Gln Gly Arg Pro Gln Pro Glu Val Thr Trp Arg Ile Asn Gly Ile
1355 1360 1365 Pro Val
Glu Glu Leu Ala Lys Asp Gln Lys Tyr Arg Ile Gln Arg 1370
1375 1380 Gly Ala Leu Ile Leu Ser Asn
Val Gln Pro Ser Asp Thr Met Val 1385 1390
1395 Thr Gln Cys Glu Ala Arg Asn Arg His Gly Leu Leu
Leu Ala Asn 1400 1405 1410
Ala Tyr Ile Tyr Val Val Gln Leu Pro Ala Lys Ile Leu Thr Ala 1415
1420 1425 Asp Asn Gln Thr Tyr
Met Ala Val Gln Gly Ser Thr Ala Tyr Leu 1430 1435
1440 Leu Cys Lys Ala Phe Gly Ala Pro Val Pro
Ser Val Gln Trp Leu 1445 1450 1455
Asp Glu Asp Gly Thr Thr Val Leu Gln Asp Glu Arg Phe Phe Pro
1460 1465 1470 Tyr Ala
Asn Gly Thr Leu Gly Ile Arg Asp Leu Gln Ala Asn Asp 1475
1480 1485 Thr Gly Arg Tyr Phe Cys Leu
Ala Ala Asn Asp Gln Asn Asn Val 1490 1495
1500 Thr Ile Met Ala Asn Leu Lys Val Lys Asp Ala Thr
Gln Ile Thr 1505 1510 1515
Gln Gly Pro Arg Ser Thr Ile Glu Lys Lys Gly Ser Arg Val Thr 1520
1525 1530 Phe Thr Cys Gln Ala
Ser Phe Asp Pro Ser Leu Gln Pro Ser Ile 1535 1540
1545 Thr Trp Arg Gly Asp Gly Arg Asp Leu Gln
Glu Leu Gly Asp Ser 1550 1555 1560
Asp Lys Tyr Phe Ile Glu Asp Gly Arg Leu Val Ile His Ser Leu
1565 1570 1575 Asp Tyr
Ser Asp Gln Gly Asn Tyr Ser Cys Val Ala Ser Thr Glu 1580
1585 1590 Leu Asp Val Val Glu Ser Arg
Ala Gln Leu Leu Val Val Gly Ser 1595 1600
1605 Pro Gly Pro Val Pro Arg Leu Val Leu Ser Asp Leu
His Leu Leu 1610 1615 1620
Thr Gln Ser Gln Val Arg Val Ser Trp Ser Pro Ala Glu Asp His 1625
1630 1635 Asn Ala Pro Ile Glu
Lys Tyr Asp Ile Glu Phe Glu Asp Lys Glu 1640 1645
1650 Met Ala Pro Glu Lys Trp Tyr Ser Leu Gly
Lys Val Pro Gly Asn 1655 1660 1665
Gln Thr Ser Thr Thr Leu Lys Leu Ser Pro Tyr Val His Tyr Thr
1670 1675 1680 Phe Arg
Val Thr Ala Ile Asn Lys Tyr Gly Pro Gly Glu Pro Ser 1685
1690 1695 Pro Val Ser Glu Thr Val Val
Thr Pro Glu Ala Ala Pro Glu Lys 1700 1705
1710 Asn Pro Val Asp Val Lys Gly Glu Gly Asn Glu Thr
Thr Asn Met 1715 1720 1725
Val Ile Thr Trp Lys Pro Leu Arg Trp Met Asp Trp Asn Ala Pro 1730
1735 1740 Gln Val Gln Tyr Arg
Val Gln Trp Arg Pro Gln Gly Thr Arg Gly 1745 1750
1755 Pro Trp Gln Glu Gln Ile Val Ser Asp Pro
Phe Leu Val Val Ser 1760 1765 1770
Asn Thr Ser Thr Phe Val Pro Tyr Glu Ile Lys Val Gln Ala Val
1775 1780 1785 Asn Ser
Gln Gly Lys Gly Pro Glu Pro Gln Val Thr Ile Gly Tyr 1790
1795 1800 Ser Gly Glu Asp Tyr Pro Gln
Ala Ile Pro Glu Leu Glu Gly Ile 1805 1810
1815 Glu Ile Leu Asn Ser Ser Ala Val Leu Val Lys Trp
Arg Pro Val 1820 1825 1830
Asp Leu Ala Gln Val Lys Gly His Leu Arg Gly Tyr Asn Val Thr 1835
1840 1845 Tyr Trp Arg Glu Gly
Ser Gln Arg Lys His Ser Lys Arg His Ile 1850 1855
1860 His Lys Asp His Val Val Val Pro Ala Asn
Thr Thr Ser Val Ile 1865 1870 1875
Leu Ser Gly Leu Arg Pro Tyr Ser Ser Tyr His Leu Glu Val Gln
1880 1885 1890 Ala Phe
Asn Gly Arg Gly Ser Gly Pro Ala Ser Glu Phe Thr Phe 1895
1900 1905 Ser Thr Pro Glu Gly Val Pro
Gly His Pro Glu Ala Leu His Leu 1910 1915
1920 Glu Cys Gln Ser Asn Thr Ser Leu Leu Leu Arg Trp
Gln Pro Pro 1925 1930 1935
Leu Ser His Asn Gly Val Leu Thr Gly Tyr Val Leu Ser Tyr His 1940
1945 1950 Pro Leu Asp Glu Gly
Gly Lys Gly Gln Leu Ser Phe Asn Leu Arg 1955 1960
1965 Asp Pro Glu Leu Arg Thr His Asn Leu Thr
Asp Leu Ser Pro His 1970 1975 1980
Leu Arg Tyr Arg Phe Gln Leu Gln Ala Thr Thr Lys Glu Gly Pro
1985 1990 1995 Gly Glu
Ala Ile Val Arg Glu Gly Gly Thr Met Ala Leu Ser Gly 2000
2005 2010 Ile Ser Asp Phe Gly Asn Ile
Ser Ala Thr Ala Gly Glu Asn Tyr 2015 2020
2025 Ser Val Val Ser Trp Val Pro Lys Glu Gly Gln Cys
Asn Phe Arg 2030 2035 2040
Phe His Ile Leu Phe Lys Ala Leu Gly Glu Glu Lys Gly Gly Ala 2045
2050 2055 Ser Leu Ser Pro Gln
Tyr Val Ser Tyr Asn Gln Ser Ser Tyr Thr 2060 2065
2070 Gln Trp Asp Leu Gln Pro Asp Thr Asp Tyr
Glu Ile His Leu Phe 2075 2080 2085
Lys Glu Arg Met Phe Arg His Gln Met Ala Val Lys Thr Asn Gly
2090 2095 2100 Thr Gly
Arg Val Arg Leu Pro Pro Ala Gly Phe Ala Thr Glu Gly 2105
2110 2115 Trp Phe Ile Gly Phe Val Ser
Ala Ile Ile Leu Leu Leu Leu Val 2120 2125
2130 Leu Leu Ile Leu Cys Phe Ile Lys Arg Ser Lys Gly
Gly Lys Tyr 2135 2140 2145
Ser Val Lys Asp Lys Glu Asp Thr Gln Val Asp Ser Glu Ala Arg 2150
2155 2160 Pro Met Lys Asp Glu
Thr Phe Gly Glu Tyr Arg Ser Leu Glu Ser 2165 2170
2175 Asp Asn Glu Glu Lys Ala Phe Gly Ser Ser
Gln Pro Ser Leu Asn 2180 2185 2190
Gly Asp Ile Lys Pro Leu Gly Ser Asp Asp Ser Leu Ala Asp Tyr
2195 2200 2205 Gly Gly
Ser Val Asp Val Gln Phe Asn Glu Asp Gly Ser Phe Ile 2210
2215 2220 Gly Gln Tyr Ser Gly Lys Lys
Glu Lys Glu Ala Ala Gly Gly Asn 2225 2230
2235 Asp Ser Ser Gly Ala Thr Ser Pro Ile Asn Pro Ala
Val Ala Leu 2240 2245 2250
Glu 342259PRTArtificialSynthetic polypeptide, C terminal fusion
chimera between extracellular domain L1 and chondroitinase ABCI
protein with a spacer or linking peptide 34Ala Thr Ser Asn Pro Ala Phe
Asp Pro Lys Asn Leu Met Gln Ser Glu 1 5
10 15 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala
Asp Phe Ser Ser Asp 20 25
30 Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly
Asn 35 40 45 Gln
Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50
55 60 Lys Lys Leu Ile Val Pro
Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly 65 70
75 80 Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu
Tyr Asn Glu Lys Pro 85 90
95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr
100 105 110 Ser Glu
Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp 115
120 125 Arg Thr Val Gly Val Ser Leu
Asn Asn Asp Leu Glu Asn Arg Glu Met 130 135
140 Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr
Gln Asp Ser Ile 145 150 155
160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro
165 170 175 Ser Asn Val
Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser 180
185 190 Val Asp Asp Ala Arg Tyr Gln Trp
Ser Asp Tyr Gln Val Lys Thr Arg 195 200
205 Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro
Gln Leu Pro 210 215 220
Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu 225
230 235 240 Ile Asn Glu Phe
Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu 245
250 255 Glu Asn Ile Ser Lys Leu Lys Ser Asp
Phe Asp Ala Leu Asn Thr His 260 265
270 Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr
Asp Lys 275 280 285
Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln 290
295 300 Leu Phe Asp Asn Tyr
Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305 310
315 320 Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys
Asp Pro Thr Gln Lys Ala 325 330
335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp
Gln 340 345 350 Gly
Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr 355
360 365 Ser Ser Arg Trp Trp Tyr
Ile Ser Thr Leu Leu Met Ser Asp Ala Leu 370 375
380 Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp
Ser Leu Leu Trp Tyr 385 390 395
400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser
405 410 415 Ser Asp
Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu 420
425 430 Leu Leu Leu Glu Pro Asp Asp
Gln Lys Arg Ile Asn Leu Val Asn Thr 435 440
445 Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val
Pro Pro Gly Gly 450 455 460
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn 465
470 475 480 Tyr Pro Gly
Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile 485
490 495 Tyr Leu Leu Arg Asp Thr Pro Phe
Ser Val Gly Glu Ser Gly Trp Asn 500 505
510 Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser
Asn Pro Glu 515 520 525
Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu 530
535 540 Lys Ser Val Ala
Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545 550
555 560 Ser Pro Asp Lys Thr Leu Ala Ser Ile
Tyr Leu Ala Ile Ser Asp Lys 565 570
575 Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr
Pro Ala 580 585 590
Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile
595 600 605 His Arg Trp Gln
Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610
615 620 Val Trp Ser Ser Glu Ile Tyr Asn
Lys Asp Asn Arg Tyr Gly Arg Tyr 625 630
635 640 Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly
Ser Gln Leu Ser 645 650
655 Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala
660 665 670 Thr Thr Ile
His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His 675
680 685 Thr Leu Met Gln Arg Gly Glu Arg
Gly Phe Ser Gly Thr Ser Ser Leu 690 695
700 Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr
Pro Ala Asn 705 710 715
720 Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala
725 730 735 Ala Asp Asn His
Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp 740
745 750 Lys Asn Lys Asn Val Glu Thr Thr Leu
Phe Gln His Ala Ile Thr Pro 755 760
765 Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn
Met Pro 770 775 780
Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785
790 795 800 Asn Gly Tyr Leu Ile
Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln 805
810 815 His Gln Val Ser Ala Glu Asn Lys Asn Arg
Gln Pro Thr Glu Gly Asn 820 825
830 Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala
Ser 835 840 845 Tyr
Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850
855 860 Met Ala Gln Lys Phe Arg
Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg 865 870
875 880 Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys
Leu Ser Asn Val Thr 885 890
895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys
900 905 910 Lys Val
Asn Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr 915
920 925 Leu Ile Val Ser Ala Val Thr
Pro Asp Leu Asn Met Thr Arg Gln Lys 930 935
940 Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn
Gly Lys Trp Gln 945 950 955
960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn
965 970 975 Thr Glu Leu
Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys 980
985 990 Leu Ser Pro Leu Pro Gly Gly Gly
Gly Gly Met Val Val Ala Leu Arg 995 1000
1005 Tyr Val Trp Pro Leu Leu Leu Cys Ser Pro Cys
Leu Leu Ile Gln 1010 1015 1020
Ile Pro Glu Glu Tyr Glu Gly His His Val Met Glu Pro Pro Val
1025 1030 1035 Ile Thr Glu
Gln Ser Pro Arg Arg Leu Val Val Phe Pro Thr Asp 1040
1045 1050 Asp Ile Ser Leu Lys Cys Glu Ala
Ser Gly Lys Pro Glu Val Gln 1055 1060
1065 Phe Arg Trp Thr Arg Asp Gly Val His Phe Lys Pro Lys
Glu Glu 1070 1075 1080
Leu Gly Val Thr Val Tyr Gln Ser Pro His Ser Gly Ser Phe Thr 1085
1090 1095 Ile Thr Gly Asn Asn
Ser Asn Phe Ala Gln Arg Phe Gln Gly Ile 1100 1105
1110 Tyr Arg Cys Phe Ala Ser Asn Lys Leu Gly
Thr Ala Met Ser His 1115 1120 1125
Glu Ile Arg Leu Met Ala Glu Gly Ala Pro Lys Trp Pro Lys Glu
1130 1135 1140 Thr Val
Lys Pro Val Glu Val Glu Glu Gly Glu Ser Val Val Leu 1145
1150 1155 Pro Cys Asn Pro Pro Pro Ser
Ala Glu Pro Leu Arg Ile Tyr Trp 1160 1165
1170 Met Asn Ser Lys Ile Leu His Ile Lys Gln Asp Glu
Arg Val Thr 1175 1180 1185
Met Gly Gln Asn Gly Asn Leu Tyr Phe Ala Asn Val Leu Thr Ser 1190
1195 1200 Asp Asn His Ser Asp
Tyr Ile Cys His Ala His Phe Pro Gly Thr 1205 1210
1215 Arg Thr Ile Ile Gln Lys Glu Pro Ile Asp
Leu Arg Val Lys Ala 1220 1225 1230
Thr Asn Ser Met Ile Asp Arg Lys Pro Arg Leu Leu Phe Pro Thr
1235 1240 1245 Asn Ser
Ser Ser His Leu Val Ala Leu Gln Gly Gln Pro Leu Val 1250
1255 1260 Leu Glu Cys Ile Ala Glu Gly
Phe Pro Thr Pro Thr Ile Lys Trp 1265 1270
1275 Leu Arg Pro Ser Gly Pro Met Pro Ala Asp Arg Val
Thr Tyr Gln 1280 1285 1290
Asn His Asn Lys Thr Leu Gln Leu Leu Lys Val Gly Glu Glu Asp 1295
1300 1305 Asp Gly Glu Tyr Arg
Cys Leu Ala Glu Asn Ser Leu Gly Ser Ala 1310 1315
1320 Arg His Ala Tyr Tyr Val Thr Val Glu Ala
Ala Pro Tyr Trp Leu 1325 1330 1335
His Lys Pro Gln Ser His Leu Tyr Gly Pro Gly Glu Thr Ala Arg
1340 1345 1350 Leu Asp
Cys Gln Val Gln Gly Arg Pro Gln Pro Glu Val Thr Trp 1355
1360 1365 Arg Ile Asn Gly Ile Pro Val
Glu Glu Leu Ala Lys Asp Gln Lys 1370 1375
1380 Tyr Arg Ile Gln Arg Gly Ala Leu Ile Leu Ser Asn
Val Gln Pro 1385 1390 1395
Ser Asp Thr Met Val Thr Gln Cys Glu Ala Arg Asn Arg His Gly 1400
1405 1410 Leu Leu Leu Ala Asn
Ala Tyr Ile Tyr Val Val Gln Leu Pro Ala 1415 1420
1425 Lys Ile Leu Thr Ala Asp Asn Gln Thr Tyr
Met Ala Val Gln Gly 1430 1435 1440
Ser Thr Ala Tyr Leu Leu Cys Lys Ala Phe Gly Ala Pro Val Pro
1445 1450 1455 Ser Val
Gln Trp Leu Asp Glu Asp Gly Thr Thr Val Leu Gln Asp 1460
1465 1470 Glu Arg Phe Phe Pro Tyr Ala
Asn Gly Thr Leu Gly Ile Arg Asp 1475 1480
1485 Leu Gln Ala Asn Asp Thr Gly Arg Tyr Phe Cys Leu
Ala Ala Asn 1490 1495 1500
Asp Gln Asn Asn Val Thr Ile Met Ala Asn Leu Lys Val Lys Asp 1505
1510 1515 Ala Thr Gln Ile Thr
Gln Gly Pro Arg Ser Thr Ile Glu Lys Lys 1520 1525
1530 Gly Ser Arg Val Thr Phe Thr Cys Gln Ala
Ser Phe Asp Pro Ser 1535 1540 1545
Leu Gln Pro Ser Ile Thr Trp Arg Gly Asp Gly Arg Asp Leu Gln
1550 1555 1560 Glu Leu
Gly Asp Ser Asp Lys Tyr Phe Ile Glu Asp Gly Arg Leu 1565
1570 1575 Val Ile His Ser Leu Asp Tyr
Ser Asp Gln Gly Asn Tyr Ser Cys 1580 1585
1590 Val Ala Ser Thr Glu Leu Asp Val Val Glu Ser Arg
Ala Gln Leu 1595 1600 1605
Leu Val Val Gly Ser Pro Gly Pro Val Pro Arg Leu Val Leu Ser 1610
1615 1620 Asp Leu His Leu Leu
Thr Gln Ser Gln Val Arg Val Ser Trp Ser 1625 1630
1635 Pro Ala Glu Asp His Asn Ala Pro Ile Glu
Lys Tyr Asp Ile Glu 1640 1645 1650
Phe Glu Asp Lys Glu Met Ala Pro Glu Lys Trp Tyr Ser Leu Gly
1655 1660 1665 Lys Val
Pro Gly Asn Gln Thr Ser Thr Thr Leu Lys Leu Ser Pro 1670
1675 1680 Tyr Val His Tyr Thr Phe Arg
Val Thr Ala Ile Asn Lys Tyr Gly 1685 1690
1695 Pro Gly Glu Pro Ser Pro Val Ser Glu Thr Val Val
Thr Pro Glu 1700 1705 1710
Ala Ala Pro Glu Lys Asn Pro Val Asp Val Lys Gly Glu Gly Asn 1715
1720 1725 Glu Thr Thr Asn Met
Val Ile Thr Trp Lys Pro Leu Arg Trp Met 1730 1735
1740 Asp Trp Asn Ala Pro Gln Val Gln Tyr Arg
Val Gln Trp Arg Pro 1745 1750 1755
Gln Gly Thr Arg Gly Pro Trp Gln Glu Gln Ile Val Ser Asp Pro
1760 1765 1770 Phe Leu
Val Val Ser Asn Thr Ser Thr Phe Val Pro Tyr Glu Ile 1775
1780 1785 Lys Val Gln Ala Val Asn Ser
Gln Gly Lys Gly Pro Glu Pro Gln 1790 1795
1800 Val Thr Ile Gly Tyr Ser Gly Glu Asp Tyr Pro Gln
Ala Ile Pro 1805 1810 1815
Glu Leu Glu Gly Ile Glu Ile Leu Asn Ser Ser Ala Val Leu Val 1820
1825 1830 Lys Trp Arg Pro Val
Asp Leu Ala Gln Val Lys Gly His Leu Arg 1835 1840
1845 Gly Tyr Asn Val Thr Tyr Trp Arg Glu Gly
Ser Gln Arg Lys His 1850 1855 1860
Ser Lys Arg His Ile His Lys Asp His Val Val Val Pro Ala Asn
1865 1870 1875 Thr Thr
Ser Val Ile Leu Ser Gly Leu Arg Pro Tyr Ser Ser Tyr 1880
1885 1890 His Leu Glu Val Gln Ala Phe
Asn Gly Arg Gly Ser Gly Pro Ala 1895 1900
1905 Ser Glu Phe Thr Phe Ser Thr Pro Glu Gly Val Pro
Gly His Pro 1910 1915 1920
Glu Ala Leu His Leu Glu Cys Gln Ser Asn Thr Ser Leu Leu Leu 1925
1930 1935 Arg Trp Gln Pro Pro
Leu Ser His Asn Gly Val Leu Thr Gly Tyr 1940 1945
1950 Val Leu Ser Tyr His Pro Leu Asp Glu Gly
Gly Lys Gly Gln Leu 1955 1960 1965
Ser Phe Asn Leu Arg Asp Pro Glu Leu Arg Thr His Asn Leu Thr
1970 1975 1980 Asp Leu
Ser Pro His Leu Arg Tyr Arg Phe Gln Leu Gln Ala Thr 1985
1990 1995 Thr Lys Glu Gly Pro Gly Glu
Ala Ile Val Arg Glu Gly Gly Thr 2000 2005
2010 Met Ala Leu Ser Gly Ile Ser Asp Phe Gly Asn Ile
Ser Ala Thr 2015 2020 2025
Ala Gly Glu Asn Tyr Ser Val Val Ser Trp Val Pro Lys Glu Gly 2030
2035 2040 Gln Cys Asn Phe Arg
Phe His Ile Leu Phe Lys Ala Leu Gly Glu 2045 2050
2055 Glu Lys Gly Gly Ala Ser Leu Ser Pro Gln
Tyr Val Ser Tyr Asn 2060 2065 2070
Gln Ser Ser Tyr Thr Gln Trp Asp Leu Gln Pro Asp Thr Asp Tyr
2075 2080 2085 Glu Ile
His Leu Phe Lys Glu Arg Met Phe Arg His Gln Met Ala 2090
2095 2100 Val Lys Thr Asn Gly Thr Gly
Arg Val Arg Leu Pro Pro Ala Gly 2105 2110
2115 Phe Ala Thr Glu Gly Trp Phe Ile Gly Phe Val Ser
Ala Ile Ile 2120 2125 2130
Leu Leu Leu Leu Val Leu Leu Ile Leu Cys Phe Ile Lys Arg Ser 2135
2140 2145 Lys Gly Gly Lys Tyr
Ser Val Lys Asp Lys Glu Asp Thr Gln Val 2150 2155
2160 Asp Ser Glu Ala Arg Pro Met Lys Asp Glu
Thr Phe Gly Glu Tyr 2165 2170 2175
Arg Ser Leu Glu Ser Asp Asn Glu Glu Lys Ala Phe Gly Ser Ser
2180 2185 2190 Gln Pro
Ser Leu Asn Gly Asp Ile Lys Pro Leu Gly Ser Asp Asp 2195
2200 2205 Ser Leu Ala Asp Tyr Gly Gly
Ser Val Asp Val Gln Phe Asn Glu 2210 2215
2220 Asp Gly Ser Phe Ile Gly Gln Tyr Ser Gly Lys Lys
Glu Lys Glu 2225 2230 2235
Ala Ala Gly Gly Asn Asp Ser Ser Gly Ala Thr Ser Pro Ile Asn 2240
2245 2250 Pro Ala Val Ala Leu
Glu 2255 35990PRTArtificialSynthetic polypeptide,
Chondroitinase ABC II protein 35Leu Pro Thr Leu Ser His Glu Ala Phe
Gly Asp Ile Tyr Leu Phe Glu 1 5 10
15 Gly Glu Leu Pro Asn Ile Leu Thr Thr Ser Asn Asn Asn Gln
Leu Ser 20 25 30
Leu Ser Lys Gln His Ala Lys Asp Gly Glu Gln Ser Leu Lys Trp Gln
35 40 45 Tyr Gln Pro Gln
Ala Thr Leu Thr Leu Asn Asn Ile Val Asn Tyr Gln 50
55 60 Asp Asp Lys Asn Thr Ala Thr Pro
Leu Thr Phe Met Met Trp Ile Tyr 65 70
75 80 Asn Glu Lys Pro Gln Ser Ser Pro Leu Thr Leu Ala
Phe Lys Gln Asn 85 90
95 Asn Lys Ile Ala Leu Ser Phe Asn Ala Glu Leu Asn Phe Thr Gly Trp
100 105 110 Arg Gly Ile
Ala Val Pro Phe Arg Asp Met Gln Gly Ser Ala Thr Gly 115
120 125 Gln Leu Asp Gln Leu Val Ile Thr
Ala Pro Asn Gln Ala Gly Thr Leu 130 135
140 Phe Phe Asp Gln Ile Ile Met Ser Val Pro Leu Asp Asn
Arg Trp Ala 145 150 155
160 Val Pro Asp Tyr Gln Thr Pro Tyr Val Asn Asn Ala Val Asn Thr Met
165 170 175 Val Ser Lys Asn
Trp Ser Ala Leu Leu Met Tyr Asp Gln Met Phe Gln 180
185 190 Ala His Tyr Pro Thr Leu Asn Phe Asp
Thr Glu Phe Arg Asp Asp Gln 195 200
205 Thr Glu Met Ala Ser Ile Tyr Gln Arg Phe Glu Tyr Tyr Gln
Gly Ile 210 215 220
Arg Ser Asp Lys Lys Ile Thr Pro Asp Met Leu Asp Lys His Leu Ala 225
230 235 240 Leu Trp Glu Lys Leu
Gly Leu Thr Gln His Ala Asp Gly Ser Ile Thr 245
250 255 Gly Lys Ala Leu Asp His Pro Asn Arg Gln
His Phe Met Lys Val Glu 260 265
270 Gly Val Phe Ser Glu Gly Thr Gln Lys Ala Leu Leu Asp Ala Asn
Met 275 280 285 Leu
Arg Asp Val Gly Lys Thr Leu Leu Gln Thr Ala Ile Tyr Leu Arg 290
295 300 Ser Asp Ser Leu Ser Ala
Thr Gly Arg Lys Lys Leu Glu Glu Arg Tyr 305 310
315 320 Leu Leu Gly Thr Arg Tyr Val Leu Glu Gln Gly
Phe Thr Arg Gly Ser 325 330
335 Gly Tyr Gln Ile Ile Thr His Val Gly Tyr Gln Thr Arg Glu Leu Phe
340 345 350 Asp Ala
Trp Phe Ile Gly Arg His Val Leu Ala Lys Asn Asn Leu Leu 355
360 365 Ala Pro Thr Gln Gln Ala Met
Met Trp Tyr Asn Ala Thr Gly Arg Ile 370 375
380 Phe Glu Lys Asp Asn Glu Ile Val Asp Ala Asn Val
Asp Ile Leu Asn 385 390 395
400 Thr Gln Leu Gln Trp Met Ile Lys Ser Leu Leu Met Leu Pro Asp Tyr
405 410 415 Gln Gln Arg
Gln Gln Ala Leu Ala Gln Leu Gln Ser Trp Leu Asn Lys 420
425 430 Thr Ile Leu Ser Ser Lys Gly Val
Ala Gly Gly Phe Lys Ser Asp Gly 435 440
445 Ser Ile Phe His His Ser Gln His Tyr Pro Ala Tyr Ala
Lys Asp Ala 450 455 460
Phe Gly Gly Leu Ala Pro Ser Val Tyr Ala Leu Ser Asp Ser Pro Phe 465
470 475 480 Arg Leu Ser Thr
Ser Ala His Glu His Leu Lys Asp Val Leu Leu Lys 485
490 495 Met Arg Ile Tyr Thr Lys Glu Thr Gln
Ile Pro Val Val Leu Ser Gly 500 505
510 Arg His Pro Thr Gly Leu His Lys Ile Gly Ile Ala Pro Phe
Lys Trp 515 520 525
Met Ala Leu Ala Gly Thr Pro Asp Gly Lys Gln Lys Leu Asp Thr Thr 530
535 540 Leu Ser Ala Ala Tyr
Ala Asn Leu Asp Asn Lys Thr His Phe Glu Gly 545 550
555 560 Ile Asn Ala Glu Ser Glu Pro Val Gly Ala
Trp Ala Met Asn Tyr Ala 565 570
575 Ser Met Ala Ile Gln Arg Arg Ala Ser Thr Gln Ser Pro Gln Gln
Ser 580 585 590 Trp
Leu Ala Ile Ala Arg Gly Phe Ser Arg Tyr Leu Val Gly Asn Glu 595
600 605 Ser Tyr Glu Asn Asn Asn
Arg Tyr Gly Arg Tyr Leu Gln Tyr Gly Gln 610 615
620 Leu Glu Ile Ile Pro Ala Asp Leu Thr Gln Ser
Gly Phe Ser His Ala 625 630 635
640 Gly Trp Asp Trp Asn Arg Tyr Pro Gly Thr Thr Thr Ile His Leu Pro
645 650 655 Tyr Asn
Glu Leu Glu Ala Lys Leu Asn Gln Leu Pro Ala Ala Gly Ile 660
665 670 Glu Glu Met Leu Leu Ser Thr
Glu Ser Tyr Ser Gly Ala Asn Thr Leu 675 680
685 Asn Asn Asn Ser Met Phe Ala Met Lys Leu His Gly
His Ser Lys Tyr 690 695 700
Gln Gln Gln Ser Leu Arg Ala Asn Lys Ser Tyr Phe Leu Phe Asp Asn 705
710 715 720 Arg Val Ile
Ala Leu Gly Ser Gly Ile Glu Asn Asp Asp Lys Gln His 725
730 735 Thr Thr Glu Thr Thr Leu Phe Gln
Phe Ala Val Pro Lys Leu Gln Ser 740 745
750 Val Ile Ile Asn Gly Lys Lys Val Asn Gln Leu Asp Thr
Gln Leu Thr 755 760 765
Leu Asn Asn Ala Asp Thr Leu Ile Asp Pro Ala Gly Asn Leu Tyr Lys 770
775 780 Leu Thr Lys Gly
Gln Thr Val Lys Phe Ser Tyr Gln Lys Gln His Ser 785 790
795 800 Leu Asp Asp Arg Asn Ser Lys Pro Thr
Glu Gln Leu Phe Ala Thr Ala 805 810
815 Val Ile Ser His Gly Lys Ala Pro Ser Asn Glu Asn Tyr Glu
Tyr Ala 820 825 830
Ile Ala Ile Glu Ala Gln Asn Asn Lys Ala Pro Lys Tyr Thr Val Leu
835 840 845 Gln His Asn Asp
Gln Leu His Ala Val Lys Asp Lys Ile Thr Gln Glu 850
855 860 Glu Gly Tyr Gly Phe Phe Glu Ala
Thr Lys Leu Lys Ser Ala Asp Ala 865 870
875 880 Thr Leu Leu Ser Ser Asp Ala Pro Val Met Val Met
Ala Lys Ile Gln 885 890
895 Asn Gln Gln Leu Thr Leu Ser Ile Val Asn Pro Asp Leu Asn Leu Tyr
900 905 910 Gln Gly Arg
Glu Lys Asp Gln Phe Asp Asp Lys Gly Asn Gln Ile Glu 915
920 925 Val Ser Val Tyr Ser Arg His Trp
Leu Thr Ala Glu Ser Gln Ser Thr 930 935
940 Asn Ser Thr Ile Thr Val Lys Gly Ile Trp Lys Leu Thr
Thr Pro Gln 945 950 955
960 Pro Gly Val Ile Ile Lys His His Asn Asn Asn Thr Leu Ile Thr Thr
965 970 975 Thr Thr Ile Gln
Ala Thr Pro Thr Val Ile Asn Leu Val Lys 980
985 990 36700PRTUnknownPedobacter heparinus
Chondroitinase AC protein 36Met Lys Lys Leu Phe Val Thr Cys Ile Val Phe
Phe Ser Ile Leu Ser 1 5 10
15 Pro Ala Leu Leu Ile Ala Gln Gln Thr Gly Thr Ala Glu Leu Ile Met
20 25 30 Lys Arg
Val Met Leu Asp Leu Lys Lys Pro Leu Arg Asn Met Asp Lys 35
40 45 Val Ala Glu Lys Asn Leu Asn Thr Leu
Gln Pro Asp Gly Ser Trp Lys 50 55 60
Asp Val Pro Tyr Lys Asp Asp Ala Met Thr Asn Trp Leu Pro Asn
Asn 65 70 75 80 His
Leu Leu Gln Leu Glu Thr Ile Ile Gln Ala Tyr Ile Glu Lys Asp
85 90 95 Ser His Tyr Tyr Gly Asp
Asp Lys Val Phe Asp Gln Ile Ser Lys Ala 100
105 110 Phe Lys Tyr Trp Tyr Asp Ser Asp Pro Lys
Ser Arg Asn Trp Trp His 115 120
125 Asn Glu Ile Ala Thr Pro Gln Ala Leu Gly Glu Met Leu Ile
Leu Met 130 135 140
Arg Tyr Gly Lys Lys Pro Leu Asp Glu Ala Leu Val His Lys Leu Thr 145
150 155 160 Glu Arg Met Lys Arg
Gly Glu Pro Glu Lys Lys Thr Gly Ala Asn Lys 165
170 175 Thr Asp Ile Ala Leu His Tyr Phe Tyr Arg
Ala Leu Leu Thr Ser Asp 180 185
190 Glu Ala Leu Leu Ser Phe Ala Val Lys Glu Leu Phe Tyr Pro Val
Gln 195 200 205 Phe
Val His Tyr Glu Glu Gly Leu Gln Tyr Asp Tyr Ser Tyr Leu Gln 210
215 220 His Gly Pro Gln Leu Gln
Ile Ser Ser Tyr Gly Ala Val Phe Ile Thr 225 230
235 240 Gly Val Leu Lys Leu Ala Asn Tyr Val Arg Asp
Thr Pro Tyr Ala Leu 245 250
255 Ser Thr Glu Lys Leu Ala Ile Phe Ser Lys Tyr Tyr Arg Asp Ser Tyr
260 265 270 Leu Lys
Ala Ile Arg Gly Ser Tyr Met Asp Phe Asn Val Glu Gly Arg 275
280 285 Gly Val Ser Arg Pro Asp Ile
Leu Asn Lys Lys Ala Glu Lys Lys Arg 290 295
300 Leu Leu Val Ala Lys Met Ile Asp Leu Lys His Thr
Glu Glu Trp Ala 305 310 315
320 Asp Ala Ile Ala Arg Thr Asp Ser Thr Val Ala Ala Gly Tyr Lys Ile
325 330 335 Glu Pro Tyr
His His Gln Phe Trp Asn Gly Asp Tyr Val Gln His Leu 340
345 350 Arg Pro Ala Tyr Ser Phe Asn Val
Arg Met Val Ser Lys Arg Thr Arg 355 360
365 Arg Ser Glu Ser Gly Asn Lys Glu Asn Leu Leu Gly Arg
Tyr Leu Ser 370 375 380
Asp Gly Ala Thr Asn Ile Gln Leu Arg Gly Pro Glu Tyr Tyr Asn Ile 385
390 395 400 Met Pro Val Trp
Glu Trp Asp Lys Ile Pro Gly Ile Thr Ser Arg Asp 405
410 415 Tyr Leu Thr Asp Arg Pro Leu Thr Lys
Leu Trp Gly Glu Gln Gly Ser 420 425
430 Asn Asp Phe Ala Gly Gly Val Ser Asp Gly Val Tyr Gly Ala
Ser Ala 435 440 445
Tyr Ala Leu Asp Tyr Asp Ser Leu Gln Ala Lys Lys Ala Trp Phe Phe 450
455 460 Phe Asp Lys Glu Ile
Val Cys Leu Gly Ala Gly Ile Asn Ser Asn Ala 465 470
475 480 Pro Glu Asn Ile Thr Thr Thr Leu Asn Gln
Ser Trp Leu Asn Gly Pro 485 490
495 Val Ile Ser Thr Ala Gly Lys Thr Gly Arg Gly Lys Ile Thr Thr
Phe 500 505 510 Lys
Ala Gln Gly Gln Phe Trp Leu Leu His Asp Ala Ile Gly Tyr Tyr 515
520 525 Phe Pro Glu Gly Ala Asn
Leu Ser Leu Ser Thr Gln Ser Gln Lys Gly 530 535
540 Asn Trp Phe His Ile Asn Asn Ser His Ser Lys
Asp Glu Val Ser Gly 545 550 555
560 Asp Val Phe Lys Leu Trp Ile Asn His Gly Ala Arg Pro Glu Asn Ala
565 570 575 Gln Tyr
Ala Tyr Ile Val Leu Pro Gly Ile Asn Lys Pro Glu Glu Ile 580
585 590 Lys Lys Tyr Asn Gly Thr Ala
Pro Lys Val Leu Ala Asn Thr Asn Gln 595 600
605 Leu Gln Ala Val Tyr His Gln Gln Leu Asp Met Val
Gln Ala Ile Phe 610 615 620
Tyr Thr Ala Gly Lys Leu Ser Val Ala Gly Ile Glu Ile Glu Thr Asp 625
630 635 640 Lys Pro Cys
Ala Val Leu Ile Lys His Ile Asn Gly Lys Gln Val Ile 645
650 655 Trp Ala Ala Asp Pro Leu Gln Lys
Glu Lys Thr Ala Val Leu Ser Ile 660 665
670 Arg Asp Leu Lys Thr Gly Lys Thr Asn Arg Val Lys Ile
Asp Phe Pro 675 680 685
Gln Gln Glu Phe Ala Gly Ala Thr Val Glu Leu Lys 690
695 700 37506PRTUnknownPedobacter heparinus
Chondroitinase B protein 37Met Lys Met Leu Asn Lys Leu Ala Gly Tyr Leu
Leu Pro Ile Met Val 1 5 10
15 Leu Leu Asn Val Ala Pro Cys Leu Gly Gln Val Val Ala Ser Asn Glu
20 25 30 Thr Leu
Tyr Gln Val Val Lys Glu Val Lys Pro Gly Gly Leu Val Gln 35
40 45 Ile Ala Asp Gly Thr Tyr Lys
Asp Val Gln Leu Ile Val Ser Asn Ser 50 55
60 Gly Lys Ser Gly Leu Pro Ile Thr Ile Lys Ala Leu
Asn Pro Gly Lys 65 70 75
80 Val Phe Phe Thr Gly Asp Ala Lys Val Glu Leu Arg Gly Glu His Leu
85 90 95 Ile Leu Glu
Gly Ile Trp Phe Lys Asp Gly Asn Arg Ala Ile Gln Ala 100
105 110 Trp Lys Ser His Gly Pro Gly Leu
Val Ala Ile Tyr Gly Ser Tyr Asn 115 120
125 Arg Ile Thr Ala Cys Val Phe Asp Cys Phe Asp Glu Ala
Asn Ser Ala 130 135 140
Tyr Ile Thr Thr Ser Leu Thr Glu Asp Gly Lys Val Pro Gln His Cys 145
150 155 160 Arg Ile Asp His
Cys Ser Phe Thr Asp Lys Ile Thr Phe Asp Gln Val 165
170 175 Ile Asn Leu Asn Asn Thr Ala Arg Ala
Ile Lys Asp Gly Ser Val Gly 180 185
190 Gly Pro Gly Met Tyr His Arg Val Asp His Cys Phe Phe Ser
Asn Pro 195 200 205
Gln Lys Pro Gly Asn Ala Gly Gly Gly Ile Arg Ile Gly Tyr Tyr Arg 210
215 220 Asn Asp Ile Gly Arg
Cys Leu Val Asp Ser Asn Leu Phe Met Arg Gln 225 230
235 240 Asp Ser Glu Ala Glu Ile Ile Thr Ser Lys
Ser Gln Glu Asn Val Tyr 245 250
255 Tyr Gly Asn Thr Tyr Leu Asn Cys Gln Gly Thr Met Asn Phe Arg
His 260 265 270 Gly
Asp His Gln Val Ala Ile Asn Asn Phe Tyr Ile Gly Asn Asp Gln 275
280 285 Arg Phe Gly Tyr Gly Gly
Met Phe Val Trp Gly Ser Arg His Val Ile 290 295
300 Ala Cys Asn Tyr Phe Glu Leu Ser Glu Thr Ile
Lys Ser Arg Gly Asn 305 310 315
320 Ala Ala Leu Tyr Leu Asn Pro Gly Ala Met Ala Ser Glu His Ala Leu
325 330 335 Ala Phe
Asp Met Leu Ile Ala Asn Asn Ala Phe Ile Asn Val Asn Gly 340
345 350 Tyr Ala Ile His Phe Asn Pro
Leu Asp Glu Arg Arg Lys Glu Tyr Cys 355 360
365 Ala Ala Asn Arg Leu Lys Phe Glu Thr Pro His Gln
Leu Met Leu Lys 370 375 380
Gly Asn Leu Phe Phe Lys Asp Lys Pro Tyr Val Tyr Pro Phe Phe Lys 385
390 395 400 Asp Asp Tyr
Phe Ile Ala Gly Lys Asn Ser Trp Thr Gly Asn Val Ala 405
410 415 Leu Gly Val Glu Lys Gly Ile Pro
Val Asn Ile Ser Ala Asn Arg Ser 420 425
430 Ala Tyr Lys Pro Val Lys Ile Lys Asp Ile Gln Pro Ile
Glu Gly Ile 435 440 445
Ala Leu Asp Leu Asn Ala Leu Ile Ser Lys Gly Ile Thr Gly Lys Pro 450
455 460 Leu Ser Trp Asp
Glu Val Arg Pro Tyr Trp Leu Lys Glu Met Pro Gly 465 470
475 480 Thr Tyr Ala Leu Thr Ala Arg Leu Ser
Ala Asp Arg Ala Ala Lys Phe 485 490
495 Lys Ala Val Ile Lys Arg Asn Lys Glu His 500
505 38405PRTUnknownHomo sapiens Hyaluronidase-1
protein 38Met Ala Ala His Leu Leu Pro Ile Cys Ala Leu Phe Leu Thr Leu Leu
1 5 10 15 Asp Met
Ala Gln Gly Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe 20
25 30 Thr Thr Val Trp Asn Ala Asn
Thr Gln Trp Cys Leu Glu Arg His Gly 35 40
45 Val Asp Val Asp Val Ser Val Phe Asp Val Val Ala
Asn Pro Gly Gln 50 55 60
Thr Phe Arg Gly Pro Asp Met Thr Ile Phe Tyr Ser Ser Gln Leu Gly 65
70 75 80 Thr Tyr Pro
Tyr Tyr Thr Pro Thr Gly Glu Pro Val Phe Gly Gly Leu 85
90 95 Pro Gln Asn Ala Ser Leu Ile Ala
His Leu Ala Arg Thr Phe Gln Asp 100 105
110 Ile Leu Ala Ala Ile Pro Ala Pro Asp Phe Ser Gly Leu
Ala Val Ile 115 120 125
Asp Trp Glu Ala Trp Arg Pro Arg Trp Ala Phe Asn Trp Asp Thr Lys 130
135 140 Asp Ile Tyr Arg
Gln Arg Ser Arg Ala Leu Val Gln Ala Gln His Pro 145 150
155 160 Asp Trp Pro Ala Pro Gln Val Glu Ala
Val Ala Gln Asp Gln Phe Gln 165 170
175 Gly Ala Ala Arg Ala Trp Met Ala Gly Thr Leu Gln Leu Gly
Arg Ala 180 185 190
Leu Arg Pro Arg Gly Leu Trp Gly Phe Tyr Gly Phe Pro Asp Cys Tyr
195 200 205 Asn Tyr Asp Phe
Leu Ser Pro Asn Tyr Thr Gly Gln Cys Pro Ser Gly 210
215 220 Ile Arg Ala Gln Asn Asp Gln Leu
Gly Trp Leu Trp Gly Gln Ser Arg 225 230
235 240 Ala Leu Tyr Pro Ser Ile Tyr Met Pro Ala Val Leu
Glu Gly Thr Gly 245 250
255 Lys Ser Gln Met Tyr Val Gln His Arg Val Ala Glu Ala Phe Arg Val
260 265 270 Ala Val Ala
Ala Gly Asp Pro Asn Leu Pro Val Leu Pro Tyr Val Gln 275
280 285 Ile Phe Tyr Asp Thr Thr Asn His
Phe Leu Pro Leu Glu Ser Cys Gln 290 295
300 Ala Ile Lys Glu Tyr Met Asp Thr Thr Leu Gly Pro Phe
Ile Leu Asn 305 310 315
320 Val Thr Ser Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Ser Gly His
325 330 335 Gly Arg Cys Val
Arg Arg Thr Ser His Pro Lys Ala Leu Leu Leu Leu 340
345 350 Asn Pro Ala Ser Phe Ser Ile Gln Leu
Thr Pro Gly Gly Gly Pro Leu 355 360
365 Ser Leu Arg Gly Ala Leu Ser Leu Glu Asp Gln Ala Gln Met
Ala Val 370 375 380
Glu Phe Lys Cys Arg Cys Tyr Pro Gly Trp Gln Ala Pro Trp Cys Glu 385
390 395 400 Arg Lys Ser Met Trp
405 39473PRTUnknownHomo sapiens Hyaluronidase-2 protein
39Met Arg Ala Gly Pro Gly Pro Thr Val Thr Leu Ala Leu Val Leu Ala 1
5 10 15 Val Ser Trp Ala
Met Glu Leu Lys Pro Thr Ala Pro Pro Ile Phe Thr 20
25 30 Gly Arg Pro Phe Val Val Ala Trp Asp
Val Pro Thr Gln Asp Cys Gly 35 40
45 Pro Arg Leu Lys Val Pro Leu Asp Leu Asn Ala Phe Asp Val
Gln Ala 50 55 60
Ser Pro Asn Glu Gly Phe Val Asn Gln Asn Ile Thr Ile Phe Tyr Arg 65
70 75 80 Asp Arg Leu Gly Leu
Tyr Pro Arg Phe Asp Ser Ala Gly Arg Ser Val 85
90 95 His Gly Gly Val Pro Gln Asn Val Ser Leu
Trp Ala His Arg Lys Met 100 105
110 Leu Gln Lys Arg Val Glu His Tyr Ile Arg Thr Gln Glu Ser Ala
Gly 115 120 125 Leu
Ala Val Ile Asp Trp Glu Asp Trp Arg Pro Val Trp Val Arg Asn 130
135 140 Trp Gln Asp Lys Asp Val
Tyr Arg Arg Leu Ser Arg Gln Leu Val Ala 145 150
155 160 Ser Arg His Pro Asp Trp Pro Pro Asp Arg Ile
Val Lys Gln Ala Gln 165 170
175 Tyr Glu Phe Glu Phe Ala Ala Gln Gln Phe Met Leu Glu Thr Leu Arg
180 185 190 Tyr Val
Lys Ala Val Arg Pro Arg His Leu Trp Gly Phe Tyr Leu Phe 195
200 205 Pro Asp Cys Tyr Asn His Asp
Tyr Val Gln Asn Trp Glu Ser Tyr Thr 210 215
220 Gly Arg Cys Pro Asp Val Glu Val Ala Arg Asn Asp
Gln Leu Ala Trp 225 230 235
240 Leu Trp Ala Glu Ser Thr Ala Leu Phe Pro Ser Val Tyr Leu Asp Glu
245 250 255 Thr Leu Ala
Ser Ser Arg His Gly Arg Asn Phe Val Ser Phe Arg Val 260
265 270 Gln Glu Ala Leu Arg Val Ala Arg
Thr His His Ala Asn His Ala Leu 275 280
285 Pro Val Tyr Val Phe Thr Arg Pro Thr Tyr Ser Arg Arg
Leu Thr Gly 290 295 300
Leu Ser Glu Met Asp Leu Ile Ser Thr Ile Gly Glu Ser Ala Ala Leu 305
310 315 320 Gly Ala Ala Gly
Val Ile Leu Trp Gly Asp Ala Gly Tyr Thr Thr Ser 325
330 335 Thr Glu Thr Cys Gln Tyr Leu Lys Asp
Tyr Leu Thr Arg Leu Leu Val 340 345
350 Pro Tyr Val Val Asn Val Ser Trp Ala Thr Gln Tyr Cys Ser
Arg Ala 355 360 365
Gln Cys His Gly His Gly Arg Cys Val Arg Arg Asn Pro Ser Ala Ser 370
375 380 Thr Phe Leu His Leu
Ser Thr Asn Ser Phe Arg Leu Val Pro Gly His 385 390
395 400 Ala Pro Gly Glu Pro Gln Leu Arg Pro Val
Gly Glu Leu Ser Trp Ala 405 410
415 Asp Ile Asp His Leu Gln Thr His Phe Arg Cys Gln Cys Tyr Leu
Gly 420 425 430 Trp
Ser Gly Glu Gln Cys Gln Trp Asp His Arg Gln Ala Ala Gly Gly 435
440 445 Ala Ser Glu Ala Trp Ala
Gly Ser His Leu Thr Ser Leu Leu Ala Leu 450 455
460 Ala Ala Leu Ala Phe Thr Trp Thr Leu 465
470 40417PRTUnknownHomo sapiens Hyaluronidase-3
protein 40Met Thr Thr Gln Leu Gly Pro Ala Leu Val Leu Gly Val Ala Leu Cys
1 5 10 15 Leu Gly
Cys Gly Gln Pro Leu Pro Gln Val Pro Glu Arg Pro Phe Ser 20
25 30 Val Leu Trp Asn Val Pro Ser
Ala His Cys Glu Ala Arg Phe Gly Val 35 40
45 His Leu Pro Leu Asn Ala Leu Gly Ile Ile Ala Asn
Arg Gly Gln His 50 55 60
Phe His Gly Gln Asn Met Thr Ile Phe Tyr Lys Asn Gln Leu Gly Leu 65
70 75 80 Tyr Pro Tyr
Phe Gly Pro Arg Gly Thr Ala His Asn Gly Gly Ile Pro 85
90 95 Gln Ala Leu Pro Leu Asp Arg His
Leu Ala Leu Ala Ala Tyr Gln Ile 100 105
110 His His Ser Leu Arg Pro Gly Phe Ala Gly Pro Ala Val
Leu Asp Trp 115 120 125
Glu Glu Trp Cys Pro Leu Trp Ala Gly Asn Trp Gly Arg Arg Arg Ala 130
135 140 Tyr Gln Ala Ala
Ser Trp Ala Trp Ala Gln Gln Val Phe Pro Asp Leu 145 150
155 160 Asp Pro Gln Glu Gln Leu Tyr Lys Ala
Tyr Thr Gly Phe Glu Gln Ala 165 170
175 Ala Arg Ala Leu Met Glu Asp Thr Leu Arg Val Ala Gln Ala
Leu Arg 180 185 190
Pro His Gly Leu Trp Gly Phe Tyr His Tyr Pro Ala Cys Gly Asn Gly
195 200 205 Trp His Ser Met
Ala Ser Asn Tyr Thr Gly Arg Cys His Ala Ala Thr 210
215 220 Leu Ala Arg Asn Thr Gln Leu His
Trp Leu Trp Ala Ala Ser Ser Ala 225 230
235 240 Leu Phe Pro Ser Ile Tyr Leu Pro Pro Arg Leu Pro
Pro Ala His His 245 250
255 Gln Ala Phe Val Arg His Arg Leu Glu Glu Ala Phe Arg Val Ala Leu
260 265 270 Val Gly His
Arg His Pro Leu Pro Val Leu Ala Tyr Val Arg Leu Thr 275
280 285 His Arg Arg Ser Gly Arg Phe Leu
Ser Gln Asp Asp Leu Val Gln Ser 290 295
300 Ile Gly Val Ser Ala Ala Leu Gly Ala Ala Gly Val Val
Leu Trp Gly 305 310 315
320 Asp Leu Ser Leu Ser Ser Ser Glu Glu Glu Cys Trp His Leu His Asp
325 330 335 Tyr Leu Val Asp
Thr Leu Gly Pro Tyr Val Ile Asn Val Thr Arg Ala 340
345 350 Ala Met Ala Cys Ser His Gln Arg Cys
His Gly His Gly Arg Cys Ala 355 360
365 Arg Arg Asp Pro Gly Gln Met Glu Ala Phe Leu His Leu Trp
Pro Asp 370 375 380
Gly Ser Leu Gly Asp Trp Lys Ser Phe Ser Cys His Cys Tyr Trp Gly 385
390 395 400 Trp Ala Gly Pro Thr
Cys Gln Glu Pro Arg Pro Gly Pro Lys Glu Ala 405
410 415 Val 41481PRTUnknownHomo sapiens
Hyaluronidase-4 protein 41Met Lys Val Leu Ser Glu Gly Gln Leu Lys Leu Cys
Val Val Gln Pro 1 5 10
15 Val His Leu Thr Ser Trp Leu Leu Ile Phe Phe Ile Leu Lys Ser Ile
20 25 30 Ser Cys Leu
Lys Pro Ala Arg Leu Pro Ile Tyr Gln Arg Lys Pro Phe 35
40 45 Ile Ala Ala Trp Asn Ala Pro Thr
Asp Gln Cys Leu Ile Lys Tyr Asn 50 55
60 Leu Arg Leu Asn Leu Lys Met Phe Pro Val Ile Gly Ser
Pro Leu Ala 65 70 75
80 Lys Ala Arg Gly Gln Asn Val Thr Ile Phe Tyr Val Asn Arg Leu Gly
85 90 95 Tyr Tyr Pro Trp
Tyr Thr Ser Gln Gly Val Pro Ile Asn Gly Gly Leu 100
105 110 Pro Gln Asn Ile Ser Leu Gln Val His
Leu Glu Lys Ala Asp Gln Asp 115 120
125 Ile Asn Tyr Tyr Ile Pro Ala Glu Asp Phe Ser Gly Leu Ala
Val Ile 130 135 140
Asp Trp Glu Tyr Trp Arg Pro Gln Trp Ala Arg Asn Trp Asn Ser Lys 145
150 155 160 Asp Val Tyr Arg Gln
Lys Ser Arg Lys Leu Ile Ser Asp Met Gly Lys 165
170 175 Asn Val Ser Ala Thr Asp Ile Glu Tyr Leu
Ala Lys Val Thr Phe Glu 180 185
190 Glu Ser Ala Lys Ala Phe Met Lys Glu Thr Ile Lys Leu Gly Ile
Lys 195 200 205 Ser
Arg Pro Lys Gly Leu Trp Gly Tyr Tyr Leu Tyr Pro Asp Cys His 210
215 220 Asn Tyr Asn Val Tyr Ala
Pro Asn Tyr Ser Gly Ser Cys Pro Glu Asp 225 230
235 240 Glu Val Leu Arg Asn Asn Glu Leu Ser Trp Leu
Trp Asn Ser Ser Ala 245 250
255 Ala Leu Tyr Pro Ser Ile Cys Val Trp Lys Ser Leu Gly Asp Ser Glu
260 265 270 Asn Ile
Leu Arg Phe Ser Lys Phe Arg Val His Glu Ser Met Arg Ile 275
280 285 Ser Thr Met Thr Ser His Asp
Tyr Ala Leu Pro Val Phe Val Tyr Thr 290 295
300 Arg Leu Gly Tyr Arg Asp Glu Pro Leu Phe Phe Leu
Ser Lys Gln Asp 305 310 315
320 Leu Val Ser Thr Ile Gly Glu Ser Ala Ala Leu Gly Ala Ala Gly Ile
325 330 335 Val Ile Trp
Gly Asp Met Asn Leu Thr Ala Ser Lys Ala Asn Cys Thr 340
345 350 Lys Val Lys Gln Phe Val Ser Ser
Asp Leu Gly Ser Tyr Ile Ala Asn 355 360
365 Val Thr Arg Ala Ala Glu Val Cys Ser Leu His Leu Cys
Arg Asn Asn 370 375 380
Gly Arg Cys Ile Arg Lys Met Trp Asn Ala Pro Ser Tyr Leu His Leu 385
390 395 400 Asn Pro Ala Ser
Tyr His Ile Glu Ala Ser Glu Asp Gly Glu Phe Thr 405
410 415 Val Lys Gly Lys Ala Ser Asp Thr Asp
Leu Ala Val Met Ala Asp Thr 420 425
430 Phe Ser Cys His Cys Tyr Gln Gly Tyr Glu Gly Ala Asp Cys
Arg Glu 435 440 445
Ile Lys Thr Ala Asp Gly Cys Ser Gly Val Ser Pro Ser Pro Gly Ser 450
455 460 Leu Met Thr Leu Cys
Leu Leu Leu Leu Ala Ser Tyr Arg Ser Ile Gln 465 470
475 480 Leu 42509PRTUnknownHomo sapiens PH-20
protein 42Met Gly Val Leu Lys Phe Lys His Ile Phe Phe Arg Ser Phe Val Lys
1 5 10 15 Ser Ser
Gly Val Ser Gln Ile Val Phe Thr Phe Leu Leu Ile Pro Cys 20
25 30 Cys Leu Thr Leu Asn Phe Arg
Ala Pro Pro Val Ile Pro Asn Val Pro 35 40
45 Phe Leu Trp Ala Trp Asn Ala Pro Ser Glu Phe Cys
Leu Gly Lys Phe 50 55 60
Asp Glu Pro Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg 65
70 75 80 Ile Asn Ala
Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu 85
90 95 Gly Tyr Tyr Pro Tyr Ile Asp Ser
Ile Thr Gly Val Thr Val Asn Gly 100 105
110 Gly Ile Pro Gln Lys Ile Ser Leu Gln Asp His Leu Asp
Lys Ala Lys 115 120 125
Lys Asp Ile Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val 130
135 140 Ile Asp Trp Glu
Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro 145 150
155 160 Lys Asp Val Tyr Lys Asn Arg Ser Ile
Glu Leu Val Gln Gln Gln Asn 165 170
175 Val Gln Leu Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln
Glu Phe 180 185 190
Glu Lys Ala Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys
195 200 205 Leu Leu Arg Pro
Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys 210
215 220 Tyr Asn His His Tyr Lys Lys Pro
Gly Tyr Asn Gly Ser Cys Phe Asn 225 230
235 240 Val Glu Ile Lys Arg Asn Asp Asp Leu Ser Trp Leu
Trp Asn Glu Ser 245 250
255 Thr Ala Leu Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val
260 265 270 Ala Ala Thr
Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val 275
280 285 Ser Lys Ile Pro Asp Ala Lys Ser
Pro Leu Pro Val Phe Ala Tyr Thr 290 295
300 Arg Ile Val Phe Thr Asp Gln Val Leu Lys Phe Leu Ser
Gln Asp Glu 305 310 315
320 Leu Val Tyr Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile
325 330 335 Val Ile Trp Gly
Thr Leu Ser Ile Met Arg Ser Met Lys Ser Cys Leu 340
345 350 Leu Leu Asp Asn Tyr Met Glu Thr Ile
Leu Asn Pro Tyr Ile Ile Asn 355 360
365 Val Thr Leu Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln
Glu Gln 370 375 380
Gly Val Cys Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu 385
390 395 400 Asn Pro Asp Asn Phe
Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr 405
410 415 Val Arg Gly Lys Pro Thr Leu Glu Asp Leu
Glu Gln Phe Ser Glu Lys 420 425
430 Phe Tyr Cys Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala
Asp 435 440 445 Val
Lys Asp Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys 450
455 460 Ile Asp Ala Phe Leu Lys
Pro Pro Met Glu Thr Glu Glu Pro Gln Ile 465 470
475 480 Phe Tyr Asn Ala Ser Pro Ser Thr Leu Ser Ala
Thr Met Phe Ile Val 485 490
495 Ser Ile Leu Phe Leu Ile Ile Ser Ser Val Ala Ser Leu
500 505 4316PRTArtificialSynthetic
polypeptide, amino acids 35-50 HIV-1 Rev protein 43Arg Gln Ala Arg
Arg Asn Arg Arg Arg Arg Trp Arg Glu Arg Gln Arg 1 5
10 15 4419PRTArtificialSynthetic
polypeptide, amino acids 1-19 of HTLV-1 Rex protein 44Met Pro Lys
Thr Arg Arg Arg Pro Arg Arg Ser Gln Arg Lys Arg Pro 1 5
10 15 Pro Thr Pro
4516PRTArtificialSynthetic polypeptide, coresponding to the third
helix of the homeodomain of Antennapedia (Derossi, et al., J. Biol.
Chem. 27118188-93, 1996) 45Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg
Met Lys Trp Lys Lys 1 5 10
15 4630PRTArtificialSynthetic polypeptide, corresponding to the
heavy chain variable region of an anti-DNA monoclonal antibody
LOCUS (AAC40104) 46Val Ala Tyr Ile Ser Arg Gly Gly Val Ser Thr Tyr Tyr
Ser Asp Thr 1 5 10 15
Val Lys Gly Arg Phe Thr Arg Gln Lys Tyr Asn Lys Arg Ala 20
25 30 47246PRTUnknownhuman herpesvirus
HSV VP22 protein 47Met Thr Ser Arg Arg Ser Val Lys Ser Gly Pro Arg Glu
Val Pro Arg 1 5 10 15
Asp Glu Tyr Glu Asp Leu Tyr Tyr Thr Pro Ser Ser Gly Met Ala Ser
20 25 30 Pro Asp Ser Pro
Pro Asp Thr Ser Arg Arg Gly Ala Leu Gln Thr Arg 35
40 45 Ser Arg Gln Arg Gly Glu Val Arg Phe
Val Gln Tyr Asp Glu Ser Asp 50 55
60 Tyr Ala Leu Tyr Gly Gly Ser Ser Ser Glu Asp Asp Glu
His Pro Glu 65 70 75
80 Val Pro Arg Thr Arg Arg Pro Val Ser Gly Ala Val Leu Ser Gly Pro
85 90 95 Gly Pro Ala Arg
Ala Pro Pro Pro Pro Ala Gly Ser Gly Gly Ala Gly 100
105 110 Arg Thr Pro Thr Thr Ala Pro Arg Ala
Pro Arg Thr Gln Arg Val Ala 115 120
125 Thr Lys Ala Pro Ala Ala Pro Ala Ala Glu Thr Thr Arg Gly
Arg Lys 130 135 140
Ser Ala Gln Pro Glu Ser Ala Ala Leu Pro Asp Ala Pro Ala Ser Arg 145
150 155 160 Ala Pro Thr Val Gln
Leu Trp Gln Met Ser Arg Pro Arg Thr Asp Glu 165
170 175 Asp Leu Asn Glu Leu Leu Gly Ile Thr His
Arg Val Thr Val Cys Glu 180 185
190 Gly Lys Asn Leu Leu Gln Arg Ala Asn Glu Leu Val Asn Pro Asp
Val 195 200 205 Val
Gln Asp Val Asp Ala Ala Thr Ala Thr Arg Gly Arg Ser Ala Ala 210
215 220 Ser Arg Pro Thr Glu Arg
Pro Arg Ala Pro Ala Arg Ser Ala Ser Arg 225 230
235 240 Pro Arg Arg Pro Val Glu 245
4836PRTArtificialSynthetic peptide, residues 37-72 of the HIV-1
Tat protein 48Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly Arg Lys Lys
Arg 1 5 10 15 Arg
Gln Arg Arg Arg Pro Pro Gln Gly Ser Gln Thr His Gln Val Ser
20 25 30 Leu Ser Lys Gln
35 4910PRTArtificialSynthetic polypeptide, amino acid residues
48-57 of (HIV-1) Tat protein 49Gly Arg Lys Lys Arg Arg Gln Arg Arg
Arg 1 5 10 50997PRTArtificialSynthetic
polypeptide, Chondroitinase ABC I protein 50Ala Thr Ser Asn Pro Ala
Phe Asp Pro Lys Asn Leu Met Gln Ser Glu 1 5
10 15 Ile Tyr His Phe Ala Gln Asn Asn Pro Leu Ala
Asp Phe Ser Ser Asp 20 25
30 Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly
Asn 35 40 45 Gln
Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50
55 60 Lys Lys Leu Ile Val Pro
Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly 65 70
75 80 Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu
Tyr Asn Glu Lys Pro 85 90
95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr
100 105 110 Ser Glu
Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp 115
120 125 Arg Thr Val Gly Val Ser Leu
Asn Asn Asp Leu Glu Asn Arg Glu Met 130 135
140 Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr
Gln Asp Ser Ile 145 150 155
160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro
165 170 175 Ser Asn Val
Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser 180
185 190 Val Asp Asp Ala Arg Tyr Gln Trp
Ser Asp Tyr Gln Val Lys Thr Arg 195 200
205 Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro
Gln Leu Pro 210 215 220
Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu 225
230 235 240 Ile Asn Glu Phe
Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu 245
250 255 Glu Asn Ile Ser Lys Leu Lys Ser Asp
Phe Asp Ala Leu Asn Thr His 260 265
270 Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr
Asp Lys 275 280 285
Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln 290
295 300 Leu Phe Asp Asn Tyr
Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305 310
315 320 Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys
Asp Pro Thr Gln Lys Ala 325 330
335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp
Gln 340 345 350 Gly
Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr 355
360 365 Ser Ser Arg Trp Trp Tyr
Ile Ser Thr Leu Leu Met Ser Asp Ala Leu 370 375
380 Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp
Ser Leu Leu Trp Tyr 385 390 395
400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser
405 410 415 Ser Asp
Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu 420
425 430 Leu Leu Leu Glu Pro Asp Asp
Gln Lys Arg Ile Asn Leu Val Asn Thr 435 440
445 Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val
Pro Pro Gly Gly 450 455 460
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn 465
470 475 480 Tyr Pro Gly
Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile 485
490 495 Tyr Leu Leu Arg Asp Thr Pro Phe
Ser Val Gly Glu Ser Gly Trp Asn 500 505
510 Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser
Asn Pro Glu 515 520 525
Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu 530
535 540 Lys Ser Val Ala
Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545 550
555 560 Ser Pro Asp Lys Thr Leu Ala Ser Ile
Tyr Leu Ala Ile Ser Asp Lys 565 570
575 Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr
Pro Ala 580 585 590
Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile
595 600 605 His Arg Trp Gln
Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610
615 620 Val Trp Ser Ser Glu Ile Tyr Asn
Lys Asp Asn Arg Tyr Gly Arg Tyr 625 630
635 640 Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly
Ser Gln Leu Ser 645 650
655 Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala
660 665 670 Thr Thr Ile
His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His 675
680 685 Thr Leu Met Gln Arg Gly Glu Arg
Gly Phe Ser Gly Thr Ser Ser Leu 690 695
700 Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile Tyr
Pro Ala Asn 705 710 715
720 Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala
725 730 735 Ala Asp Asn His
Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp 740
745 750 Lys Asn Lys Asn Val Glu Thr Thr Leu
Phe Gln His Ala Ile Thr Pro 755 760
765 Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn
Met Pro 770 775 780
Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785
790 795 800 Asn Gly Tyr Leu Ile
Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln 805
810 815 His Gln Val Ser Ala Glu Asn Lys Asn Arg
Gln Pro Thr Glu Gly Asn 820 825
830 Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala
Ser 835 840 845 Tyr
Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850
855 860 Met Ala Gln Lys Phe Arg
Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg 865 870
875 880 Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys
Leu Ser Asn Val Thr 885 890
895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys
900 905 910 Lys Val
Asn Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr 915
920 925 Leu Ile Val Ser Ala Val Thr
Pro Asp Leu Asn Met Thr Arg Gln Lys 930 935
940 Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn
Gly Lys Trp Gln 945 950 955
960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn
965 970 975 Thr Glu Leu
Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys 980
985 990 Leu Ser Pro Leu Pro 995
51977PRTArtificialSynthetic polypeptide, N(delta)20 ABCI
(A(sub)45-N(sub)1023) protein 51Ala Gln Asn Asn Pro Leu Ala Asp Phe Ser
Ser Asp Lys Asn Ser Ile 1 5 10
15 Leu Thr Leu Ser Asp Lys Arg Ser Ile Met Gly Asn Gln Ser Leu
Leu 20 25 30 Trp
Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His Lys Lys Leu Ile 35
40 45 Val Pro Thr Asp Lys Glu
Ala Ser Lys Ala Trp Gly Arg Ser Ser Thr 50 55
60 Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu Lys
Pro Ile Asp Gly Tyr 65 70 75
80 Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala Gln
85 90 95 Ala Gly
Phe Lys Val Lys Leu Asp Phe Thr Gly Trp Arg Thr Val Gly 100
105 110 Val Ser Leu Asn Asn Asp Leu
Glu Asn Arg Glu Met Thr Leu Asn Ala 115 120
125 Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp Ser Ile
Gly Arg Ser Leu 130 135 140
Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn Val Ser 145
150 155 160 Gln Gly Glu
Ile Tyr Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala 165
170 175 Arg Tyr Gln Trp Ser Asp Tyr Gln
Val Lys Thr Arg Leu Ser Glu Pro 180 185
190 Glu Ile Gln Phe His Asn Val Lys Pro Gln Leu Pro Val
Thr Pro Glu 195 200 205
Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu Phe 210
215 220 Val Gly Gly Glu
Lys Glu Thr Asn Leu Ala Leu Glu Glu Asn Ile Ser 225 230
235 240 Lys Leu Lys Ser Asp Phe Asp Ala Leu
Asn Thr His Thr Leu Ala Asn 245 250
255 Gly Gly Thr Gln Gly Arg His Leu Ile Thr Asp Lys Gln Ile
Ile Ile 260 265 270
Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp Asn
275 280 285 Tyr Val Ile Leu
Gly Asn Tyr Thr Thr Leu Met Phe Asn Ile Ser Arg 290
295 300 Ala Tyr Val Leu Glu Lys Asp Pro
Thr Gln Lys Ala Gln Leu Lys Gln 305 310
315 320 Met Tyr Leu Leu Met Thr Lys His Leu Leu Asp Gln
Gly Phe Val Lys 325 330
335 Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr Ser Ser Arg Trp
340 345 350 Trp Tyr Ile
Ser Thr Leu Leu Met Ser Asp Ala Leu Lys Glu Ala Asn 355
360 365 Leu Gln Thr Gln Val Tyr Asp Ser
Leu Leu Trp Tyr Ser Arg Glu Phe 370 375
380 Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp Ser Ser
Asp Leu Asp 385 390 395
400 Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu
405 410 415 Pro Asp Asp Gln
Lys Arg Ile Asn Leu Val Asn Thr Phe Ser His Tyr 420
425 430 Ile Thr Gly Ala Leu Thr Gln Val Pro
Pro Gly Gly Lys Asp Gly Leu 435 440
445 Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn Tyr Pro
Gly Tyr 450 455 460
Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg 465
470 475 480 Asp Thr Pro Phe Ser
Val Gly Glu Ser Gly Trp Asn Asn Leu Lys Lys 485
490 495 Ala Met Val Ser Ala Trp Ile Tyr Ser Asn
Pro Glu Val Gly Leu Pro 500 505
510 Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val
Ala 515 520 525 Gln
Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser Ser Pro Asp Lys 530
535 540 Thr Leu Ala Ser Ile Tyr
Leu Ala Ile Ser Asp Lys Thr Gln Asn Glu 545 550
555 560 Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr Pro
Ala Ser Leu Pro Gln 565 570
575 Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln
580 585 590 Asp Lys
Met Val Thr Leu Lys Ala Tyr Asn Thr Asn Val Trp Ser Ser 595
600 605 Glu Ile Tyr Asn Lys Asp Asn
Arg Tyr Gly Arg Tyr Gln Ser His Gly 610 615
620 Val Ala Gln Ile Val Ser Asn Gly Ser Gln Leu Ser
Gln Gly Tyr Gln 625 630 635
640 Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile His
645 650 655 Leu Pro Leu
Lys Asp Leu Asp Ser Pro Lys Pro His Thr Leu Met Gln 660
665 670 Arg Gly Glu Arg Gly Phe Ser Gly
Thr Ser Ser Leu Glu Gly Gln Tyr 675 680
685 Gly Met Met Ala Phe Asn Leu Ile Tyr Pro Ala Asn Leu
Glu Arg Phe 690 695 700
Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His 705
710 715 720 Leu Ile Phe Ile
Gly Ser Asn Ile Asn Ser Ser Asp Lys Asn Lys Asn 725
730 735 Val Glu Thr Thr Leu Phe Gln His Ala
Ile Thr Pro Thr Leu Asn Thr 740 745
750 Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln
Thr Thr 755 760 765
Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu 770
775 780 Ile Thr Gln Ala Glu
Lys Val Asn Val Ser Arg Gln His Gln Val Ser 785 790
795 800 Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu
Gly Asn Phe Ser Ser Ala 805 810
815 Trp Ile Asp His Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr
Met 820 825 830 Val
Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys 835
840 845 Phe Arg Glu Asn Asn Gly
Leu Tyr Gln Val Leu Arg Lys Asp Lys Asp 850 855
860 Val His Ile Ile Leu Asp Lys Leu Ser Asn Val
Thr Gly Tyr Ala Phe 865 870 875
880 Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys Lys Val Asn Lys
885 890 895 Pro Ala
Ile Val Met Thr His Arg Gln Lys Asp Thr Leu Ile Val Ser 900
905 910 Ala Val Thr Pro Asp Leu Asn
Met Thr Arg Gln Lys Ala Ala Thr Pro 915 920
925 Val Thr Ile Asn Val Thr Ile Asn Gly Lys Trp Gln
Ser Ala Asp Lys 930 935 940
Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn Thr Glu Leu Thr 945
950 955 960 Phe Thr Ser
Tyr Phe Gly Ile Pro Gln Glu Ile Lys Leu Ser Pro Leu 965
970 975 Pro 52937PRTArtificialSynthetic
polypeptide, N(delta)60 ABCI (F(sub)85-N(sub)1023) protein 52Phe Thr
Leu His Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala Ser 1 5
10 15 Lys Ala Trp Gly Arg Ser Ser
Thr Pro Val Phe Ser Phe Trp Leu Tyr 20 25
30 Asn Glu Lys Pro Ile Asp Gly Tyr Leu Thr Ile Asp
Phe Gly Glu Lys 35 40 45
Leu Ile Ser Thr Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp
50 55 60 Phe Thr Gly
Trp Arg Thr Val Gly Val Ser Leu Asn Asn Asp Leu Glu 65
70 75 80 Asn Arg Glu Met Thr Leu Asn
Ala Thr Asn Thr Ser Ser Asp Gly Thr 85
90 95 Gln Asp Ser Ile Gly Arg Ser Leu Gly Ala Lys
Val Asp Ser Ile Arg 100 105
110 Phe Lys Ala Pro Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp
Arg 115 120 125 Ile
Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp Tyr Gln 130
135 140 Val Lys Thr Arg Leu Ser
Glu Pro Glu Ile Gln Phe His Asn Val Lys 145 150
155 160 Pro Gln Leu Pro Val Thr Pro Glu Asn Leu Ala
Ala Ile Asp Leu Ile 165 170
175 Arg Gln Arg Leu Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn
180 185 190 Leu Ala
Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp Ala 195
200 205 Leu Asn Thr His Thr Leu Ala
Asn Gly Gly Thr Gln Gly Arg His Leu 210 215
220 Ile Thr Asp Lys Gln Ile Ile Ile Tyr Gln Pro Glu
Asn Leu Asn Ser 225 230 235
240 Gln Asp Lys Gln Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr
245 250 255 Thr Leu Met
Phe Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro 260
265 270 Thr Gln Lys Ala Gln Leu Lys Gln
Met Tyr Leu Leu Met Thr Lys His 275 280
285 Leu Leu Asp Gln Gly Phe Val Lys Gly Ser Ala Leu Val
Thr Thr His 290 295 300
His Trp Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met 305
310 315 320 Ser Asp Ala Leu
Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser 325
330 335 Leu Leu Trp Tyr Ser Arg Glu Phe Lys
Ser Ser Phe Asp Met Lys Val 340 345
350 Ser Ala Asp Ser Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser
Arg Gln 355 360 365
His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln Lys Arg Ile Asn 370
375 380 Leu Val Asn Thr Phe
Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val 385 390
395 400 Pro Pro Gly Gly Lys Asp Gly Leu Arg Pro
Asp Gly Thr Ala Trp Arg 405 410
415 His Glu Gly Asn Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn
Ala 420 425 430 Ser
Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val Gly Glu 435
440 445 Ser Gly Trp Asn Asn Leu
Lys Lys Ala Met Val Ser Ala Trp Ile Tyr 450 455
460 Ser Asn Pro Glu Val Gly Leu Pro Leu Ala Gly
Arg His Pro Phe Asn 465 470 475
480 Ser Pro Ser Leu Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met
485 490 495 Ser Ala
Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu Ala 500
505 510 Ile Ser Asp Lys Thr Gln Asn
Glu Ser Thr Ala Ile Phe Gly Glu Thr 515 520
525 Ile Thr Pro Ala Ser Leu Pro Gln Gly Phe Tyr Ala
Phe Asn Gly Gly 530 535 540
Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala 545
550 555 560 Tyr Asn Thr
Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg 565
570 575 Tyr Gly Arg Tyr Gln Ser His Gly
Val Ala Gln Ile Val Ser Asn Gly 580 585
590 Ser Gln Leu Ser Gln Gly Tyr Gln Gln Glu Gly Trp Asp
Trp Asn Arg 595 600 605
Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys Asp Leu Asp Ser 610
615 620 Pro Lys Pro His
Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly 625 630
635 640 Thr Ser Ser Leu Glu Gly Gln Tyr Gly
Met Met Ala Phe Asn Leu Ile 645 650
655 Tyr Pro Ala Asn Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala
Lys Lys 660 665 670
Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile Gly Ser Asn Ile
675 680 685 Asn Ser Ser Asp
Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His 690
695 700 Ala Ile Thr Pro Thr Leu Asn Thr
Leu Trp Ile Asn Gly Gln Lys Ile 705 710
715 720 Glu Asn Met Pro Tyr Gln Thr Thr Leu Gln Gln Gly
Asp Trp Leu Ile 725 730
735 Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys Val Asn
740 745 750 Val Ser Arg
Gln His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro 755
760 765 Thr Glu Gly Asn Phe Ser Ser Ala
Trp Ile Asp His Ser Thr Arg Pro 770 775
780 Lys Asp Ala Ser Tyr Glu Tyr Met Val Phe Leu Asp Ala
Thr Pro Glu 785 790 795
800 Lys Met Gly Glu Met Ala Gln Lys Phe Arg Glu Asn Asn Gly Leu Tyr
805 810 815 Gln Val Leu Arg
Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu 820
825 830 Ser Asn Val Thr Gly Tyr Ala Phe Tyr
Gln Pro Ala Ser Ile Glu Asp 835 840
845 Lys Trp Ile Lys Lys Val Asn Lys Pro Ala Ile Val Met Thr
His Arg 850 855 860
Gln Lys Asp Thr Leu Ile Val Ser Ala Val Thr Pro Asp Leu Asn Met 865
870 875 880 Thr Arg Gln Lys Ala
Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn 885
890 895 Gly Lys Trp Gln Ser Ala Asp Lys Asn Ser
Glu Val Lys Tyr Gln Val 900 905
910 Ser Gly Asp Asn Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile
Pro 915 920 925 Gln
Glu Ile Lys Leu Ser Pro Leu Pro 930 935
53858PRTArtificialSynthetic polypeptide, N(delta) 60 C(delta)80 ABCI
(F[sub]85 - A[sub]942) 53Phe Thr Leu His Lys Lys Leu Ile Val Pro Thr Asp
Lys Glu Ala Ser 1 5 10
15 Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr
20 25 30 Asn Glu Lys
Pro Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys 35
40 45 Leu Ile Ser Thr Ser Glu Ala Gln
Ala Gly Phe Lys Val Lys Leu Asp 50 55
60 Phe Thr Gly Trp Arg Thr Val Gly Val Ser Leu Asn Asn
Asp Leu Glu 65 70 75
80 Asn Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr
85 90 95 Gln Asp Ser Ile
Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg 100
105 110 Phe Lys Ala Pro Ser Asn Val Ser Gln
Gly Glu Ile Tyr Ile Asp Arg 115 120
125 Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp
Tyr Gln 130 135 140
Val Lys Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys 145
150 155 160 Pro Gln Leu Pro Val
Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile 165
170 175 Arg Gln Arg Leu Ile Asn Glu Phe Val Gly
Gly Glu Lys Glu Thr Asn 180 185
190 Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp
Ala 195 200 205 Leu
Asn Thr His Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu 210
215 220 Ile Thr Asp Lys Gln Ile
Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser 225 230
235 240 Gln Asp Lys Gln Leu Phe Asp Asn Tyr Val Ile
Leu Gly Asn Tyr Thr 245 250
255 Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro
260 265 270 Thr Gln
Lys Ala Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His 275
280 285 Leu Leu Asp Gln Gly Phe Val
Lys Gly Ser Ala Leu Val Thr Thr His 290 295
300 His Trp Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser
Thr Leu Leu Met 305 310 315
320 Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser
325 330 335 Leu Leu Trp
Tyr Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val 340
345 350 Ser Ala Asp Ser Ser Asp Leu Asp
Tyr Phe Asn Thr Leu Ser Arg Gln 355 360
365 His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln Lys
Arg Ile Asn 370 375 380
Leu Val Asn Thr Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val 385
390 395 400 Pro Pro Gly Gly
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg 405
410 415 His Glu Gly Asn Tyr Pro Gly Tyr Ser
Phe Pro Ala Phe Lys Asn Ala 420 425
430 Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val
Gly Glu 435 440 445
Ser Gly Trp Asn Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr 450
455 460 Ser Asn Pro Glu Val
Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn 465 470
475 480 Ser Pro Ser Leu Lys Ser Val Ala Gln Gly
Tyr Tyr Trp Leu Ala Met 485 490
495 Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu
Ala 500 505 510 Ile
Ser Asp Lys Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr 515
520 525 Ile Thr Pro Ala Ser Leu
Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly 530 535
540 Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met
Val Thr Leu Lys Ala 545 550 555
560 Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg
565 570 575 Tyr Gly
Arg Tyr Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly 580
585 590 Ser Gln Leu Ser Gln Gly Tyr
Gln Gln Glu Gly Trp Asp Trp Asn Arg 595 600
605 Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys
Asp Leu Asp Ser 610 615 620
Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly 625
630 635 640 Thr Ser Ser
Leu Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile 645
650 655 Tyr Pro Ala Asn Leu Glu Arg Phe
Asp Pro Asn Phe Thr Ala Lys Lys 660 665
670 Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile Gly
Ser Asn Ile 675 680 685
Asn Ser Ser Asp Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His 690
695 700 Ala Ile Thr Pro
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile 705 710
715 720 Glu Asn Met Pro Tyr Gln Thr Thr Leu
Gln Gln Gly Asp Trp Leu Ile 725 730
735 Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys
Val Asn 740 745 750
Val Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro
755 760 765 Thr Glu Gly Asn
Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro 770
775 780 Lys Asp Ala Ser Tyr Glu Tyr Met
Val Phe Leu Asp Ala Thr Pro Glu 785 790
795 800 Lys Met Gly Glu Met Ala Gln Lys Phe Arg Glu Asn
Asn Gly Leu Tyr 805 810
815 Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu
820 825 830 Ser Asn Val
Thr Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp 835
840 845 Lys Trp Ile Lys Lys Val Asn Lys
Pro Ala 850 855
541021PRTArtificialSynthetic polypeptide, ABCI a site-specific
mutant designated as H501A and Y508A where amino acid H at position
501 in ABCI is replaced by A, and amino acid Y at position 508 in
ABCI is replaced by A. 54Met Pro Ile Phe Arg Phe Thr Ala Leu Ala Met Thr
Leu Gly Leu Leu 1 5 10
15 Ser Ala Pro Tyr Asn Ala Met Ala Ala Thr Ser Asn Pro Ala Phe Asp
20 25 30 Pro Lys Asn
Leu Met Gln Ser Glu Ile Tyr His Phe Ala Gln Asn Asn 35
40 45 Pro Leu Ala Asp Phe Ser Ser Asp
Lys Asn Ser Ile Leu Thr Leu Ser 50 55
60 Asp Lys Arg Ser Ile Met Gly Asn Gln Ser Leu Leu Trp
Lys Trp Lys 65 70 75
80 Gly Gly Ser Ser Phe Thr Leu His Lys Lys Leu Ile Val Pro Thr Asp
85 90 95 Lys Glu Ala Ser
Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser 100
105 110 Phe Trp Leu Tyr Asn Glu Lys Pro Ile
Asp Gly Tyr Leu Thr Ile Asp 115 120
125 Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala Gln Ala Gly
Phe Lys 130 135 140
Val Lys Leu Asp Phe Thr Gly Trp Arg Ala Val Gly Val Ser Leu Asn 145
150 155 160 Asn Asp Leu Glu Asn
Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser 165
170 175 Ser Asp Gly Thr Gln Asp Ser Ile Gly Arg
Ser Leu Gly Ala Lys Val 180 185
190 Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn Val Ser Gln Gly Glu
Ile 195 200 205 Tyr
Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp 210
215 220 Ser Asp Tyr Gln Val Lys
Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe 225 230
235 240 His Asn Val Lys Pro Gln Leu Pro Val Thr Pro
Glu Asn Leu Ala Ala 245 250
255 Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu Phe Val Gly Gly Glu
260 265 270 Lys Glu
Thr Asn Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser 275
280 285 Asp Phe Asp Ala Leu Asn Ile
His Thr Leu Ala Asn Gly Gly Thr Gln 290 295
300 Gly Arg His Leu Ile Thr Asp Lys Gln Ile Ile Ile
Tyr Gln Pro Glu 305 310 315
320 Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp Asn Tyr Val Ile Leu
325 330 335 Gly Asn Tyr
Thr Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val Leu 340
345 350 Glu Lys Asp Pro Thr Gln Lys Ala
Gln Leu Lys Gln Met Tyr Leu Leu 355 360
365 Met Thr Lys His Leu Leu Asp Gln Gly Phe Val Lys Gly
Ser Ala Leu 370 375 380
Val Thr Thr His His Trp Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser 385
390 395 400 Thr Leu Leu Met
Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln 405
410 415 Val Tyr Asp Ser Leu Leu Trp Tyr Ser
Arg Glu Phe Lys Ser Ser Phe 420 425
430 Asp Met Lys Val Ser Ala Asp Ser Ser Asp Leu Asp Tyr Phe
Asn Thr 435 440 445
Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln 450
455 460 Lys Arg Ile Asn Leu
Val Asn Thr Phe Ser His Tyr Ile Thr Gly Ala 465 470
475 480 Leu Thr Gln Val Pro Pro Gly Gly Lys Asp
Gly Leu Arg Pro Asp Gly 485 490
495 Thr Ala Trp Arg Ala Glu Gly Asn Tyr Pro Gly Ala Ser Phe Pro
Ala 500 505 510 Phe
Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe 515
520 525 Ser Val Gly Glu Ser Gly
Trp Asn Asn Leu Lys Lys Ala Met Val Ser 530 535
540 Ala Trp Ile Tyr Ser Asn Pro Glu Val Gly Leu
Pro Leu Ala Gly Arg 545 550 555
560 His Pro Phe Asn Ser Pro Ser Leu Lys Ser Val Ala Gln Gly Tyr Tyr
565 570 575 Trp Leu
Ala Met Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser 580
585 590 Ile Tyr Leu Ala Ile Ser Asp
Lys Thr Gln Asn Glu Ser Thr Ala Ile 595 600
605 Phe Gly Glu Thr Ile Thr Pro Ala Ser Leu Pro Gln
Gly Phe Tyr Ala 610 615 620
Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met Val 625
630 635 640 Thr Leu Lys
Ala Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn 645
650 655 Lys Asp Asn Arg Tyr Gly Arg Tyr
Gln Ser His Gly Val Ala Gln Ile 660 665
670 Val Ser Asn Gly Ser Gln Leu Ser Gln Gly Tyr Gln Gln
Glu Gly Trp 675 680 685
Asp Trp Asn Arg Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys 690
695 700 Asp Leu Asp Ser
Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg 705 710
715 720 Gly Phe Ser Gly Thr Ser Ser Leu Glu
Gly Gln Tyr Gly Met Met Ala 725 730
735 Phe Asn Leu Ile Tyr Pro Ala Asn Leu Glu Arg Phe Asp Pro
Asn Phe 740 745 750
Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile
755 760 765 Gly Ser Asn Ile
Asn Ser Ser Asp Lys Asn Lys Asn Val Glu Thr Thr 770
775 780 Leu Phe Gln His Ala Ile Thr Pro
Thr Leu Asn Thr Leu Trp Ile Asn 785 790
795 800 Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln Thr Thr
Leu Gln Gln Gly 805 810
815 Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala
820 825 830 Glu Lys Val
Asn Val Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys 835
840 845 Asn Arg Gln Pro Thr Glu Gly Asn
Phe Ser Ser Ala Trp Ile Asp His 850 855
860 Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr Met Val
Phe Leu Asp 865 870 875
880 Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys Phe Arg Glu Asn
885 890 895 Asn Gly Leu Tyr
Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile 900
905 910 Leu Asp Lys Leu Ser Asn Val Thr Gly
Tyr Ala Phe Tyr Gln Pro Ala 915 920
925 Ser Ile Glu Asp Lys Trp Ile Lys Lys Val Asn Lys Pro Ala
Ile Val 930 935 940
Met Thr His Arg Gln Lys Asp Thr Leu Ile Val Ser Ala Val Thr Pro 945
950 955 960 Asp Leu Asn Met Thr
Arg Gln Lys Ala Ala Thr Pro Val Thr Ile Asn 965
970 975 Val Thr Ile Asn Gly Lys Trp Gln Ser Ala
Asp Lys Asn Ser Glu Val 980 985
990 Lys Tyr Gln Val Ser Gly Asp Asn Thr Glu Leu Thr Phe Thr
Ser Tyr 995 1000 1005
Phe Gly Ile Pro Gln Glu Ile Lys Leu Ser Pro Leu Pro 1010
1015 1020 5540PRTArtificialSynthetic
polypeptide corresponding to amino acids 821-862 from Nogo-A protein
short form (homo sapiens) LOCUS (AAG40878) 55Arg Ile Tyr Lys Gly Val
Ile Gln Ala Ile Gln Lys Ser Asp Glu Gly 1 5
10 15 His Pro Phe Arg Ala Tyr Leu Glu Ser Glu Val
Ala Ile Ser Glu Glu 20 25
30 Leu Val Gln Lys Tyr Ser Asn Ser 35
40 565PRTArtificialSynthetic polypeptide, Gly penta-linker 56Gly Gly Gly
Gly Gly 1 5 575PRTArtificialSynthetic polypeptide,
Gly-Ala linker 57Gly Gly Ala Gly Gly 1 5
582994DNAArtificialSynthetic polynucleotide for Chondroitinase ABCI
58gccaccagca atcctgcatt tgatcctaaa aatctgatgc agtcagaaat ttaccatttt
60gcacaaaata acccattagc agacttctca tcagataaaa actcaatact aacgttatct
120gataaacgta gcattatggg aaaccaatct cttttatgga aatggaaagg tggtagtagc
180tttactttac ataaaaaact gattgtcccc accgataaag aagcatctaa agcatgggga
240cgctcatcca cccccgtttt ctcattttgg ctttacaatg aaaaaccgat tgatggttat
300cttactatcg atttcggaga aaaactcatt tcaaccagtg aggctcaggc aggctttaaa
360gtaaaattag atttcactgg ctggcgtact gtgggagtct ctttaaataa cgatcttgaa
420aatcgagaga tgaccttaaa tgcaaccaat acctcctctg atggtactca agacagcatt
480gggcgttctt taggtgctaa agtcgatagt attcgtttta aagcgccttc taatgtgagt
540cagggtgaaa tctatatcga ccgtattatg ttttctgtcg atgatgctcg ctaccaatgg
600tctgattatc aagtaaaaac tcgcttatca gaacctgaaa ttcaatttca caacgtaaag
660ccacaactac ctgtaacacc tgaaaattta gcggccattg atcttattcg ccaacgtcta
720attaatgaat ttgtcggagg tgaaaaagag acaaacctcg cattagaaga gaatatcagc
780aaattaaaaa gtgatttcga tgctcttaat actcacactt tagcaaatgg tggaacgcaa
840ggcagacatc tgatcactga taaacaaatc attatttatc aaccagagaa tcttaactct
900caagataaac aactatttga taattatgtt attttaggta attacacgac attaatgttt
960aatattagcc gtgcttatgt gctggaaaaa gatcccacac aaaaggcgca actaaagcag
1020atgtacttat taatgacaaa gcatttatta gatcaaggct ttgttaaagg gagtgcttta
1080gtgacaaccc atcactgggg atacagttct cgttggtggt atatttccac gttattaatg
1140tctgatgcac taaaagaagc gaacctacaa actcaagttt atgattcatt actgtggtat
1200tcacgtgagt ttaaaagtag ttttgatatg aaagtaagtg ctgatagctc tgatctagat
1260tatttcaata ccttatctcg ccaacattta gccttattac tactagagcc tgatgatcaa
1320aagcgtatca acttagttaa tactttcagc cattatatca ctggcgcatt aacgcaagtg
1380ccaccgggtg gtaaagatgg tttacgccct gatggtacag catggcgaca tgaaggcaac
1440tatccgggct actctttccc agcctttaaa aatgcctctc agcttattta tttattacgc
1500gatacaccat tttcagtggg tgaaagtggt tggaatagcc tgaaaaaagc gatggtttca
1560gcgtggatct acagtaatcc agaagttgga ttaccgcttg caggaagaca ccctcttaac
1620tcaccttcgt taaaatcagt cgctcaaggc tattactggc ttgccatgtc tgcaaaatca
1680tcgcctgata aaacacttgc atctatttat cttgcgatta gtgataaaac acaaaatgaa
1740tcaactgcta tttttggaga aactattaca ccagcgtctt tacctcaagg tttctatgcc
1800tttaatggcg gtgcttttgg tattcatcgt tggcaagata aaatggtgac actgaaagct
1860tataacacca atgtttggtc atctgaaatt tataacaaag ataaccgtta tggccgttac
1920caaagtcatg gtgtcgctca aatagtgagt aatggctcgc agctttcaca gggctatcag
1980caagaaggtt gggattggaa tagaatgcca ggggcaacca ctatccacct tcctcttaaa
2040gacttagaca gtcctaaacc tcatacctta atgcaacgtg gagagcgtgg atttagcgga
2100acatcatccc ttgaaggtca atatggcatg atggcattcg atcttattta tcccgccaat
2160cttgagcgtt ttgatcctaa tttcactgcg aaaaagagtg tattagccgc tgataatcac
2220ttaattttta ttggtagcaa tataaatagt agtgataaaa ataaaaatgt tgaaacgacc
2280ttattccaac atgccattac tccaacatta aatacccttt ggattaatgg acaaaagata
2340gaaaacatgc cttatcaaac aacacttcaa caaggtgatt ggttaattga tagcaatggc
2400aatggttact taattactca agcagaaaaa gtaaatgtaa gtcgccaaca tcaggtttca
2460gcggaaaata aaaatcgcca accgacagaa ggaaacttta gctcggcatg gatcgatcac
2520agcactcgcc ccaaagatgc cagttatgag tatatggtct ttttagatgc gacacctgaa
2580aaaatgggag agatggcaca aaaattccgt gaaaataatg ggttatatca ggttcttcgt
2640aaggataaag acgttcatat tattctcgat aaactcagca atgtaacggg atatgccttt
2700tatcagccag catcaattga agacaaatgg atcaaaaagg ttaataaacc tgcaattgtg
2760atgactcatc gacaaaaaga cactcttatt gtcagtgcag ttacacctga tttaaatatg
2820actcgccaaa aagcagcaac tcctgtcacc atcaatgtca cgattaatgg caaatggcaa
2880tctgctgata aaaatagtga agtgaaatat caggtttctg gtgataacac tgaactgacg
2940tttacgagtt actttggtat tccacaagaa atcaaactct cgccactccc ttga
2994591924DNAUnknownHomo sapiens NgR DNA fragment corresponding to
residues 1-359 59ctgtgcgccc tgcgcgccct gcgcacccgc ggcccgagcc cagccagagc
cgggcggagc 60ggagcgcgcc gagcctcgtc ccgcggccgg gccggggccg ggccgtagcg
gcggcgcctg 120gatgcggacc cggccgcggg gagacgggcg cccgccccga aacgactttc
agtccccgac 180gcgccccgcc caacccctac gatgaagagg gcgtccgctg gagggagccg
gctgctggca 240tgggtgctgt ggctgcaggc ctggcaggtg gcagccccat gcccaggtgc
ctgcgtatgc 300tacaatgagc ccaaggtgac gacaagctgc ccccagcagg gcctgcaggc
tgtgcccgtg 360ggcatccctg ctgccagcca gcgcatcttc ctgcacggca accgcatctc
gcatgtgcca 420gctgccagct tccgtgcctg ccgcaacctc accatcctgt ggctgcactc
gaatgtgctg 480gcccgaattg atgcggctgc cttcactggc ctggccctcc tggagcagct
ggacctcagc 540gataatgcac agctccggtc tgtggaccct gccacattcc acggcctggg
ccgcctacac 600acgctgcacc tggaccgctg cggcctgcag gagctgggcc cggggctgtt
ccgcggcctg 660gctgccctgc agtacctcta cctgcaggac aacgcgctgc aggcactgcc
tgatgacacc 720ttccgcgacc tgggcaacct cacacacctc ttcctgcacg gcaaccgcat
ctccagcgtg 780cccgagcgcg ccttccgtgg gctgcacagc ctcgaccgtc tcctactgca
ccagaaccgc 840gtggcccatg tgcacccgca tgccttccgt gaccttggcc gcctcatgac
actctatctg 900tttgccaaca atctatcagc gctgcccact gaggccctgg cccccctgcg
tgccctgcag 960tacctgaggc tcaacgacaa cccctgggtg tgtgactgcc gggcacgccc
actctgggcc 1020tggctgcaga agttccgcgg ctcctcctcc gaggtgccct gcagcctccc
gcaacgcctg 1080gctggccgtg acctcaaacg cctagctgcc aatgacctgc agggctgcgc
tgtggccacc 1140ggcccttacc atcccatctg gaccggcagg gccaccgatg aggagccgct
ggggcttccc 1200aagtgctgcc agccagatgc cgctgacaag gcctcagtac tggagcctgg
aagaccagct 1260tcggcaggca atgcgctgaa gggacgcgtg ccgcccggtg acagcccgcc
gggcaacggc 1320tctggcccac ggcacatcaa tgactcaccc tttgggactc tgcctggctc
tgctgagccc 1380ccgctcactg cagtgcggcc cgagggctcc gagccaccag ggttccccac
ctcgggccct 1440cgccggaggc caggctgttc acgcaagaac cgcacccgca gccactgccg
tctgggccag 1500gcaggcagcg ggggtggcgg gactggtgac tcagaaggct caggtgccct
acccagcctc 1560acctgcagcc tcacccccct gggcctggcg ctggtgctgt ggacagtgct
tgggccctgc 1620tgacccccag cggacacaag agcgtgctca gcagccaggt gtgtgtacat
acggggtctc 1680tctccacgcc gccaagccag ccgggcggcc gacccgtggg gcaggccagg
ccaggtcctc 1740cctgatggac gcctgccgcc cgccaccccc atctccaccc catcatgttt
acagggttcg 1800gcggcagcgt ttgttccaga acgccgcctc ccacccagat cgcggtatat
agagatatgc 1860attttatttt acttgtgtaa aaatatcgga cgacgtggaa taaagagctc
ttttcttaaa 1920aaaa
192460855DNAArtificialNgR DNA fragment corresponding to
residues 27-311 60tgcccaggtg cctgcgtatg ctacaatgag cccaaggtga
cgacaagctg cccccagcag 60ggcctgcagg ctgtgcccgt gggcatccct gctgccagcc
agcgcatctt cctgcacggc 120aaccgcatct cgcatgtgcc agctgccagc ttccgtgcct
gccgcaacct caccatcctg 180tggctgcact cgaatgtgct ggcccgaatt gatgcggctg
ccttcactgg cctggccctc 240ctggagcagc tggacctcag cgataatgca cagctccggt
ctgtggaccc tgccacattc 300cacggcctgg gccgcctaca cacgctgcac ctggaccgct
gcggcctgca ggagctgggc 360ccggggctgt tccgcggcct ggctgccctg cagtacctct
acctgcagga caacgcgctg 420caggcactgc ctgatgacac cttccgcgac ctgggcaacc
tcacacacct cttcctgcac 480ggcaaccgca tctccagcgt gcccgagcgc gccttccgtg
ggctgcacag cctcgaccgt 540ctcctactgc accagaaccg cgtggcccat gtgcacccgc
atgccttccg tgaccttggc 600cgcctcatga cactctatct gtttgccaac aatctatcag
cgctgcccac tgaggccctg 660gcccccctgc gtgccctgca gtacctgagg ctcaacgaca
acccctgggt gtgtgactgc 720cgggcacgcc cactctgggc ctggctgcag aagttccgcg
gctcctcctc cgaggtgccc 780tgcagcctcc cgcaacgcct ggctggccgt gacctcaaac
gcctagctgc caatgacctg 840cagggctgcg ctgtg
855613835DNAArtificialSynthetic polynucleotide,
TAT fusion chondroitinase ABCI nucleic acid 61ggtcgtaaaa agcgtcgtca
acgtcgtcgt ggtggtggtg gtggtgccac cagcaatcct 60gcatttgatc ctaaaaatct
gatgcagtca gaaatttacc attttgcaca aaataaccca 120ttagcagact tctcatcaga
taaaaactca atactaacgt tatctgataa acgtagcatt 180atgggaaacc aatctctttt
atggaaatgg aaaggtggta gtagctttac tttacataaa 240aaactgattg tccccaccga
taaagaagca tctaaagcat ggggacgctc atctaccccc 300gttttctcat tttggcttta
caatgaaaaa ccgattgatg gttatcttac tatcgatttc 360ggagaaaaac tcatttcaac
cagtgaggct caggcaggct ttaaagtaaa attagatttc 420actggctggc gtgctgtggg
agtctcttta aataacgatc ttgaaaatcg agagatgacc 480ttaaatgcaa ccaatacctc
ctctgatggt actcaagaca gcattgggcg ttctttaggt 540gctaaagtcg atagtattcg
ttttaaagcg ccttctaatg tgagtcaggg tgaaatctat 600atcgaccgta ttatgttttc
tgtcgatgat gctcgctacc aatggtctga ttatcaagta 660aaaactcgct tatcagaacc
tgaaattcaa tttcacaacg taaagccaca actacctgta 720acacctgaaa atttagcggc
cattgatctt attcgccaac gtctaattaa tgaatttgtc 780ggaggtgaaa aagagacaaa
cctcgcatta gaagagaata tcagcaaatt aaaaagtgat 840ttcgatgctc ttaatattca
cactttagca aatggtggaa cgcaaggcag acatctgatc 900actgataaac aaatcattat
ttatcaacca gagaatctta actcccaaga taaacaacta 960tttgataatt atgttatttt
aggtaattac acgacattaa tgtttaatat tagccgtgct 1020tatgtgctgg aaaaagatcc
cacacaaaag gcgcaactaa agcagatgta cttattaatg 1080acaaagcatt tattagatca
aggctttgtt aaagggagtg ctttagtgac aacccatcac 1140tggggataca gttctcgttg
gtggtatatt tccacgttat taatgtctga tgcactaaaa 1200gaagcgaacc tacaaactca
agtttatgat tcattactgt ggtattcacg tgagtttaaa 1260agtagttttg atatgaaagt
aagtgctgat agctctgatc tagattattt caatacctta 1320tctcgccaac atttagcctt
attattacta gagcctgatg atcaaaagcg tatcaactta 1380gttaatactt tcagccatta
tatcactggc gcattaacgc aagtgccacc gggtggtaaa 1440gatggtttac gccctgatgg
tacagcatgg cgacatgaag gcaactatcc gggctactct 1500ttcccagcct ttaaaaatgc
ctctcagctt atttatttat tacgcgatac accattttca 1560gtgggtgaaa gtggttggaa
taacctgaaa aaagcgatgg tttcagcgtg gatctacagt 1620aatccagaag ttggattacc
gcttgcagga agacaccctt ttaactcacc ttcgttaaaa 1680tcagtcgctc aaggctatta
ctggcttgcc atgtctgcaa aatcatcgcc tgataaaaca 1740cttgcatcta tttatcttgc
gattagtgat aaaacacaaa atgaatcaac tgctattttt 1800ggagaaacta ttacaccagc
gtctttacct caaggtttct atgcctttaa tggcggtgct 1860tttggtattc atcgttggca
agataaaatg gtgacactga aagcttataa caccaatgtt 1920tggtcatctg aaatttataa
caaagataac cgttatggcc gttaccaaag tcatggtgtc 1980gctcaaatag tgagtaatgg
ctcgcagctt tcacagggct atcagcaaga aggttgggat 2040tggaatagaa tgcaaggggc
aaccactatt caccttcctc ttaaagactt agacagtcct 2100aaacctcata ccttaatgca
acgtggagag cgtggattta gcggaacatc atcccttgaa 2160ggtcaatatg gcatgatggc
attcgatctt atttatcccg ccaatcttga gcgttttgat 2220cctaatttca ctgcgaaaaa
gagtgtatta gccgctgata atcacttaat ttttattggt 2280agcaatataa atagtagtga
taaaaataaa aatgttgaaa cgaccttatt ccaacatgcc 2340attactccaa cattaaatac
cctttggatt aatggacaaa agatagaaaa catgccttat 2400caaacaacac ttcaacaagg
tgattggtta attgatagca atggcaatgg ttacttaatt 2460actcaagcag aaaaagtaaa
tgtaagtcgc caacatcagg tttcagcgga aaataaaaat 2520cgccaaccga cagaaggaaa
ctttagctcg gcatggatcg atcacagcac tcgccccaaa 2580gatgccagtt atgagtatat
ggtcttttta gatgcgacac ctgaaaaaat gggagagatg 2640gcacaaaaat tccgtgaaaa
taatgggtta tatcaggttc ttcgtaagga taaagacgtt 2700catattattc tcgataaact
cagcaatgta acgggatatg ccttttatca gccagcatca 2760attgaagaca aatggatcaa
aaaggttaat aaacctgcaa ttgtgatgac tcatcgacaa 2820aaagacactc ttattgtcag
tgcagttaca cctgatttaa atatgactcg ccaaaaagca 2880gcaactcctg tcaccatcaa
tgtcacgatt aatggcaaat ggcaatctgc tgataaaaat 2940agtgaagtga aatatcaggt
ttctggtgat aacactgaac tgacgtttac gagttacttt 3000ggtattccac aagaaatcaa
actctcgcca ctcccttgat ttaatcaaaa gaacgctctt 3060gcgttccttt tttatttgca
ggaaatctga ttatgctaat aaaaaaccct ttagcccacg 3120cggttacatt aagcctctgt
ttatcattac ccgcacaagc attacccact ctgtctcatg 3180aagctttcgg cgatatttat
ctttttgaag gtgaattacc caataccctt accacttcaa 3240ataataatca attatcgcta
agcaaacagc atgctaaaga tggtgaacaa tcactcaaat 3300ggcaatatca accacaagca
acattaacac taaataatat tgttaattac caagatgata 3360aaaatacagc cacaccactc
acttttatga tgtggattta taatgaaaaa cctcaatctt 3420ccccattaac gttagcattt
aaacaaaata ataaaattgc actaagtttt aatgctgaac 3480ttaattttac ggggtggcga
ggtattgctg ttccttttcg tgatatgcaa ggctctgcga 3540caggtcaact tgatcaatta
gtgatcaccg ctccaaacca agccggaaca ctcttttttg 3600atcaaatcat catgagtgta
ccgttagaca atcgttgggc agtacctgac tatcaaacac 3660cttacgtaaa taacgcagta
aacacgatgg ttagtaaaaa ctggagtgca ttattgatgt 3720acgatcagat gtttcaagcc
cattacccta ctttaaactt cgatactgaa tttcgcgatg 3780accaaacaga aatggcttcg
atttatcagc gctttgaata ttatcaagga attcc
383562478PRTArtificialSynthetic polypeptide C(delta)200
chondroitinase AC (Q[sub]23 - T[sub]500) 62Gln Gln Thr Gly Thr Ala Glu
Leu Ile Met Lys Arg Val Met Leu Asp 1 5
10 15 Leu Lys Lys Pro Leu Arg Asn Met Asp Lys Val
Ala Glu Lys Asn Leu 20 25
30 Asn Thr Leu Gln Pro Asp Gly Ser Trp Lys Asp Val Pro Tyr Lys
Asp 35 40 45 Asp
Ala Met Thr Asn Trp Leu Pro Asn Asn His Leu Leu Gln Leu Glu 50
55 60 Thr Ile Ile Gln Ala Tyr
Ile Glu Lys Asp Ser His Tyr Tyr Gly Asp 65 70
75 80 Asp Lys Val Phe Asp Gln Ile Ser Lys Ala Phe
Lys Tyr Trp Tyr Asp 85 90
95 Ser Asp Pro Lys Ser Arg Asn Trp Trp His Asn Glu Ile Ala Thr Pro
100 105 110 Gln Ala
Leu Gly Glu Met Leu Ile Leu Met Arg Tyr Gly Lys Lys Pro 115
120 125 Leu Asp Glu Ala Leu Val His
Lys Leu Thr Glu Arg Met Lys Arg Gly 130 135
140 Glu Pro Glu Lys Lys Thr Gly Ala Asn Lys Thr Asp
Ile Ala Leu His 145 150 155
160 Tyr Phe Tyr Arg Ala Leu Leu Thr Ser Asp Glu Ala Leu Leu Ser Phe
165 170 175 Ala Val Lys
Glu Leu Phe Tyr Pro Val Gln Phe Val His Tyr Glu Glu 180
185 190 Gly Leu Gln Tyr Asp Tyr Ser Tyr
Leu Gln His Gly Pro Gln Leu Gln 195 200
205 Ile Ser Ser Tyr Gly Ala Val Phe Ile Thr Gly Val Leu
Lys Leu Ala 210 215 220
Asn Tyr Val Arg Asp Thr Pro Tyr Ala Leu Ser Thr Glu Lys Leu Ala 225
230 235 240 Ile Phe Ser Lys
Tyr Tyr Arg Asp Ser Tyr Leu Lys Ala Ile Arg Gly 245
250 255 Ser Tyr Met Asp Phe Asn Val Glu Gly
Arg Gly Val Ser Arg Pro Asp 260 265
270 Ile Leu Asn Lys Lys Ala Glu Lys Lys Arg Leu Leu Val Ala
Lys Met 275 280 285
Ile Asp Leu Lys His Thr Glu Glu Trp Ala Asp Ala Ile Ala Arg Thr 290
295 300 Asp Ser Thr Val Ala
Ala Gly Tyr Lys Ile Glu Pro Tyr His His Gln 305 310
315 320 Phe Trp Asn Gly Asp Tyr Val Gln His Leu
Arg Pro Ala Tyr Ser Phe 325 330
335 Asn Val Arg Met Val Ser Lys Arg Thr Arg Arg Ser Glu Ser Gly
Asn 340 345 350 Lys
Glu Asn Leu Leu Gly Arg Tyr Leu Ser Asp Gly Ala Thr Asn Ile 355
360 365 Gln Leu Arg Gly Pro Glu
Tyr Tyr Asn Ile Met Pro Val Trp Glu Trp 370 375
380 Asp Lys Ile Pro Gly Ile Thr Ser Arg Asp Tyr
Leu Thr Asp Arg Pro 385 390 395
400 Leu Thr Lys Leu Trp Gly Glu Gln Gly Ser Asn Asp Phe Ala Gly Gly
405 410 415 Val Ser
Asp Gly Val Tyr Gly Ala Ser Ala Tyr Ala Leu Asp Tyr Asp 420
425 430 Ser Leu Gln Ala Lys Lys Ala
Trp Phe Phe Phe Asp Lys Glu Ile Val 435 440
445 Cys Leu Gly Ala Gly Ile Asn Ser Asn Ala Pro Glu
Asn Ile Thr Thr 450 455 460
Thr Leu Asn Gln Ser Trp Leu Asn Gly Pro Val Ile Ser Thr 465
470 475 63458PRTArtificialSynthetic
polypeptide C(delta)220 chondroitinase AC (Q[sub]23 - A[sub]480)
63Gln Gln Thr Gly Thr Ala Glu Leu Ile Met Lys Arg Val Met Leu Asp 1
5 10 15 Leu Lys Lys Pro
Leu Arg Asn Met Asp Lys Val Ala Glu Lys Asn Leu 20
25 30 Asn Thr Leu Gln Pro Asp Gly Ser Trp
Lys Asp Val Pro Tyr Lys Asp 35 40
45 Asp Ala Met Thr Asn Trp Leu Pro Asn Asn His Leu Leu Gln
Leu Glu 50 55 60
Thr Ile Ile Gln Ala Tyr Ile Glu Lys Asp Ser His Tyr Tyr Gly Asp 65
70 75 80 Asp Lys Val Phe Asp
Gln Ile Ser Lys Ala Phe Lys Tyr Trp Tyr Asp 85
90 95 Ser Asp Pro Lys Ser Arg Asn Trp Trp His
Asn Glu Ile Ala Thr Pro 100 105
110 Gln Ala Leu Gly Glu Met Leu Ile Leu Met Arg Tyr Gly Lys Lys
Pro 115 120 125 Leu
Asp Glu Ala Leu Val His Lys Leu Thr Glu Arg Met Lys Arg Gly 130
135 140 Glu Pro Glu Lys Lys Thr
Gly Ala Asn Lys Thr Asp Ile Ala Leu His 145 150
155 160 Tyr Phe Tyr Arg Ala Leu Leu Thr Ser Asp Glu
Ala Leu Leu Ser Phe 165 170
175 Ala Val Lys Glu Leu Phe Tyr Pro Val Gln Phe Val His Tyr Glu Glu
180 185 190 Gly Leu
Gln Tyr Asp Tyr Ser Tyr Leu Gln His Gly Pro Gln Leu Gln 195
200 205 Ile Ser Ser Tyr Gly Ala Val
Phe Ile Thr Gly Val Leu Lys Leu Ala 210 215
220 Asn Tyr Val Arg Asp Thr Pro Tyr Ala Leu Ser Thr
Glu Lys Leu Ala 225 230 235
240 Ile Phe Ser Lys Tyr Tyr Arg Asp Ser Tyr Leu Lys Ala Ile Arg Gly
245 250 255 Ser Tyr Met
Asp Phe Asn Val Glu Gly Arg Gly Val Ser Arg Pro Asp 260
265 270 Ile Leu Asn Lys Lys Ala Glu Lys
Lys Arg Leu Leu Val Ala Lys Met 275 280
285 Ile Asp Leu Lys His Thr Glu Glu Trp Ala Asp Ala Ile
Ala Arg Thr 290 295 300
Asp Ser Thr Val Ala Ala Gly Tyr Lys Ile Glu Pro Tyr His His Gln 305
310 315 320 Phe Trp Asn Gly
Asp Tyr Val Gln His Leu Arg Pro Ala Tyr Ser Phe 325
330 335 Asn Val Arg Met Val Ser Lys Arg Thr
Arg Arg Ser Glu Ser Gly Asn 340 345
350 Lys Glu Asn Leu Leu Gly Arg Tyr Leu Ser Asp Gly Ala Thr
Asn Ile 355 360 365
Gln Leu Arg Gly Pro Glu Tyr Tyr Asn Ile Met Pro Val Trp Glu Trp 370
375 380 Asp Lys Ile Pro Gly
Ile Thr Ser Arg Asp Tyr Leu Thr Asp Arg Pro 385 390
395 400 Leu Thr Lys Leu Trp Gly Glu Gln Gly Ser
Asn Asp Phe Ala Gly Gly 405 410
415 Val Ser Asp Gly Val Tyr Gly Ala Ser Ala Tyr Ala Leu Asp Tyr
Asp 420 425 430 Ser
Leu Gln Ala Lys Lys Ala Trp Phe Phe Phe Asp Lys Glu Ile Val 435
440 445 Cys Leu Gly Ala Gly Ile
Asn Ser Asn Ala 450 455
64458PRTArtificialSynthetic polypeptide, N(delta)20 C(delta)200
chondroitinase AC (L[sub]43 -T[sub]500) 64Leu Arg Asn Met Asp Lys Val Ala
Glu Lys Asn Leu Asn Thr Leu Gln 1 5 10
15 Pro Asp Gly Ser Trp Lys Asp Val Pro Tyr Lys Asp Asp
Ala Met Thr 20 25 30
Asn Trp Leu Pro Asn Asn His Leu Leu Gln Leu Glu Thr Ile Ile Gln
35 40 45 Ala Tyr Ile Glu
Lys Asp Ser His Tyr Tyr Gly Asp Asp Lys Val Phe 50
55 60 Asp Gln Ile Ser Lys Ala Phe Lys
Tyr Trp Tyr Asp Ser Asp Pro Lys 65 70
75 80 Ser Arg Asn Trp Trp His Asn Glu Ile Ala Thr Pro
Gln Ala Leu Gly 85 90
95 Glu Met Leu Ile Leu Met Arg Tyr Gly Lys Lys Pro Leu Asp Glu Ala
100 105 110 Leu Val His
Lys Leu Thr Glu Arg Met Lys Arg Gly Glu Pro Glu Lys 115
120 125 Lys Thr Gly Ala Asn Lys Thr Asp
Ile Ala Leu His Tyr Phe Tyr Arg 130 135
140 Ala Leu Leu Thr Ser Asp Glu Ala Leu Leu Ser Phe Ala
Val Lys Glu 145 150 155
160 Leu Phe Tyr Pro Val Gln Phe Val His Tyr Glu Glu Gly Leu Gln Tyr
165 170 175 Asp Tyr Ser Tyr
Leu Gln His Gly Pro Gln Leu Gln Ile Ser Ser Tyr 180
185 190 Gly Ala Val Phe Ile Thr Gly Val Leu
Lys Leu Ala Asn Tyr Val Arg 195 200
205 Asp Thr Pro Tyr Ala Leu Ser Thr Glu Lys Leu Ala Ile Phe
Ser Lys 210 215 220
Tyr Tyr Arg Asp Ser Tyr Leu Lys Ala Ile Arg Gly Ser Tyr Met Asp 225
230 235 240 Phe Asn Val Glu Gly
Arg Gly Val Ser Arg Pro Asp Ile Leu Asn Lys 245
250 255 Lys Ala Glu Lys Lys Arg Leu Leu Val Ala
Lys Met Ile Asp Leu Lys 260 265
270 His Thr Glu Glu Trp Ala Asp Ala Ile Ala Arg Thr Asp Ser Thr
Val 275 280 285 Ala
Ala Gly Tyr Lys Ile Glu Pro Tyr His His Gln Phe Trp Asn Gly 290
295 300 Asp Tyr Val Gln His Leu
Arg Pro Ala Tyr Ser Phe Asn Val Arg Met 305 310
315 320 Val Ser Lys Arg Thr Arg Arg Ser Glu Ser Gly
Asn Lys Glu Asn Leu 325 330
335 Leu Gly Arg Tyr Leu Ser Asp Gly Ala Thr Asn Ile Gln Leu Arg Gly
340 345 350 Pro Glu
Tyr Tyr Asn Ile Met Pro Val Trp Glu Trp Asp Lys Ile Pro 355
360 365 Gly Ile Thr Ser Arg Asp Tyr
Leu Thr Asp Arg Pro Leu Thr Lys Leu 370 375
380 Trp Gly Glu Gln Gly Ser Asn Asp Phe Ala Gly Gly
Val Ser Asp Gly 385 390 395
400 Val Tyr Gly Ala Ser Ala Tyr Ala Leu Asp Tyr Asp Ser Leu Gln Ala
405 410 415 Lys Lys Ala
Trp Phe Phe Phe Asp Lys Glu Ile Val Cys Leu Gly Ala 420
425 430 Gly Ile Asn Ser Asn Ala Pro Glu
Asn Ile Thr Thr Thr Leu Asn Gln 435 440
445 Ser Trp Leu Asn Gly Pro Val Ile Ser Thr 450
455 65427PRTArtificialSynthetic polypeptide,
N(delta)50 C(delta)200 of chondroitinase AC (T[sub]74 -T[sub]500)
65Thr Asn Trp Leu Pro Asn Asn His Leu Leu Gln Leu Glu Thr Ile Ile 1
5 10 15 Gln Ala Tyr Ile
Glu Lys Asp Ser His Tyr Tyr Gly Asp Asp Lys Val 20
25 30 Phe Asp Gln Ile Ser Lys Ala Phe Lys
Tyr Trp Tyr Asp Ser Asp Pro 35 40
45 Lys Ser Arg Asn Trp Trp His Asn Glu Ile Ala Thr Pro Gln
Ala Leu 50 55 60
Gly Glu Met Leu Ile Leu Met Arg Tyr Gly Lys Lys Pro Leu Asp Glu 65
70 75 80 Ala Leu Val His Lys
Leu Thr Glu Arg Met Lys Arg Gly Glu Pro Glu 85
90 95 Lys Lys Thr Gly Ala Asn Lys Thr Asp Ile
Ala Leu His Tyr Phe Tyr 100 105
110 Arg Ala Leu Leu Thr Ser Asp Glu Ala Leu Leu Ser Phe Ala Val
Lys 115 120 125 Glu
Leu Phe Tyr Pro Val Gln Phe Val His Tyr Glu Glu Gly Leu Gln 130
135 140 Tyr Asp Tyr Ser Tyr Leu
Gln His Gly Pro Gln Leu Gln Ile Ser Ser 145 150
155 160 Tyr Gly Ala Val Phe Ile Thr Gly Val Leu Lys
Leu Ala Asn Tyr Val 165 170
175 Arg Asp Thr Pro Tyr Ala Leu Ser Thr Glu Lys Leu Ala Ile Phe Ser
180 185 190 Lys Tyr
Tyr Arg Asp Ser Tyr Leu Lys Ala Ile Arg Gly Ser Tyr Met 195
200 205 Asp Phe Asn Val Glu Gly Arg
Gly Val Ser Arg Pro Asp Ile Leu Asn 210 215
220 Lys Lys Ala Glu Lys Lys Arg Leu Leu Val Ala Lys
Met Ile Asp Leu 225 230 235
240 Lys His Thr Glu Glu Trp Ala Asp Ala Ile Ala Arg Thr Asp Ser Thr
245 250 255 Val Ala Ala
Gly Tyr Lys Ile Glu Pro Tyr His His Gln Phe Trp Asn 260
265 270 Gly Asp Tyr Val Gln His Leu Arg
Pro Ala Tyr Ser Phe Asn Val Arg 275 280
285 Met Val Ser Lys Arg Thr Arg Arg Ser Glu Ser Gly Asn
Lys Glu Asn 290 295 300
Leu Leu Gly Arg Tyr Leu Ser Asp Gly Ala Thr Asn Ile Gln Leu Arg 305
310 315 320 Gly Pro Glu Tyr
Tyr Asn Ile Met Pro Val Trp Glu Trp Asp Lys Ile 325
330 335 Pro Gly Ile Thr Ser Arg Asp Tyr Leu
Thr Asp Arg Pro Leu Thr Lys 340 345
350 Leu Trp Gly Glu Gln Gly Ser Asn Asp Phe Ala Gly Gly Val
Ser Asp 355 360 365
Gly Val Tyr Gly Ala Ser Ala Tyr Ala Leu Asp Tyr Asp Ser Leu Gln 370
375 380 Ala Lys Lys Ala Trp
Phe Phe Phe Asp Lys Glu Ile Val Cys Leu Gly 385 390
395 400 Ala Gly Ile Asn Ser Asn Ala Pro Glu Asn
Ile Thr Thr Thr Leu Asn 405 410
415 Gln Ser Trp Leu Asn Gly Pro Val Ile Ser Thr 420
425 66378PRTArtificialSynthetic polypeptide,
N(delta)100 C(delta)200 of chondroitinase AC (S[sub]123 - T[sub] 500
66Ser Arg Asn Trp Trp His Asn Glu Ile Ala Thr Pro Gln Ala Leu Gly 1
5 10 15 Glu Met Leu Ile
Leu Met Arg Tyr Gly Lys Lys Pro Leu Asp Glu Ala 20
25 30 Leu Val His Lys Leu Thr Glu Arg Met
Lys Arg Gly Glu Pro Glu Lys 35 40
45 Lys Thr Gly Ala Asn Lys Thr Asp Ile Ala Leu His Tyr Phe
Tyr Arg 50 55 60
Ala Leu Leu Thr Ser Asp Glu Ala Leu Leu Ser Phe Ala Val Lys Glu 65
70 75 80 Leu Phe Tyr Pro Val
Gln Phe Val His Tyr Glu Glu Gly Leu Gln Tyr 85
90 95 Asp Tyr Ser Tyr Leu Gln His Gly Pro Gln
Leu Gln Ile Ser Ser Tyr 100 105
110 Gly Ala Val Phe Ile Thr Gly Val Leu Lys Leu Ala Asn Tyr Val
Arg 115 120 125 Asp
Thr Pro Tyr Ala Leu Ser Thr Glu Lys Leu Ala Ile Phe Ser Lys 130
135 140 Tyr Tyr Arg Asp Ser Tyr
Leu Lys Ala Ile Arg Gly Ser Tyr Met Asp 145 150
155 160 Phe Asn Val Glu Gly Arg Gly Val Ser Arg Pro
Asp Ile Leu Asn Lys 165 170
175 Lys Ala Glu Lys Lys Arg Leu Leu Val Ala Lys Met Ile Asp Leu Lys
180 185 190 His Thr
Glu Glu Trp Ala Asp Ala Ile Ala Arg Thr Asp Ser Thr Val 195
200 205 Ala Ala Gly Tyr Lys Ile Glu
Pro Tyr His His Gln Phe Trp Asn Gly 210 215
220 Asp Tyr Val Gln His Leu Arg Pro Ala Tyr Ser Phe
Asn Val Arg Met 225 230 235
240 Val Ser Lys Arg Thr Arg Arg Ser Glu Ser Gly Asn Lys Glu Asn Leu
245 250 255 Leu Gly Arg
Tyr Leu Ser Asp Gly Ala Thr Asn Ile Gln Leu Arg Gly 260
265 270 Pro Glu Tyr Tyr Asn Ile Met Pro
Val Trp Glu Trp Asp Lys Ile Pro 275 280
285 Gly Ile Thr Ser Arg Asp Tyr Leu Thr Asp Arg Pro Leu
Thr Lys Leu 290 295 300
Trp Gly Glu Gln Gly Ser Asn Asp Phe Ala Gly Gly Val Ser Asp Gly 305
310 315 320 Val Tyr Gly Ala
Ser Ala Tyr Ala Leu Asp Tyr Asp Ser Leu Gln Ala 325
330 335 Lys Lys Ala Trp Phe Phe Phe Asp Lys
Glu Ile Val Cys Leu Gly Ala 340 345
350 Gly Ile Asn Ser Asn Ala Pro Glu Asn Ile Thr Thr Thr Leu
Asn Gln 355 360 365
Ser Trp Leu Asn Gly Pro Val Ile Ser Thr 370 375
67353PRTArtificialSynthetic polypeptide, N(delta)50 C(delta)275
of chondroitinase AC (T[sub]74 - L[sub]426) 67Thr Asn Trp Leu Pro Asn
Asn His Leu Leu Gln Leu Glu Thr Ile Ile 1 5
10 15 Gln Ala Tyr Ile Glu Lys Asp Ser His Tyr Tyr
Gly Asp Asp Lys Val 20 25
30 Phe Asp Gln Ile Ser Lys Ala Phe Lys Tyr Trp Tyr Asp Ser Asp
Pro 35 40 45 Lys
Ser Arg Asn Trp Trp His Asn Glu Ile Ala Thr Pro Gln Ala Leu 50
55 60 Gly Glu Met Leu Ile Leu
Met Arg Tyr Gly Lys Lys Pro Leu Asp Glu 65 70
75 80 Ala Leu Val His Lys Leu Thr Glu Arg Met Lys
Arg Gly Glu Pro Glu 85 90
95 Lys Lys Thr Gly Ala Asn Lys Thr Asp Ile Ala Leu His Tyr Phe Tyr
100 105 110 Arg Ala
Leu Leu Thr Ser Asp Glu Ala Leu Leu Ser Phe Ala Val Lys 115
120 125 Glu Leu Phe Tyr Pro Val Gln
Phe Val His Tyr Glu Glu Gly Leu Gln 130 135
140 Tyr Asp Tyr Ser Tyr Leu Gln His Gly Pro Gln Leu
Gln Ile Ser Ser 145 150 155
160 Tyr Gly Ala Val Phe Ile Thr Gly Val Leu Lys Leu Ala Asn Tyr Val
165 170 175 Arg Asp Thr
Pro Tyr Ala Leu Ser Thr Glu Lys Leu Ala Ile Phe Ser 180
185 190 Lys Tyr Tyr Arg Asp Ser Tyr Leu
Lys Ala Ile Arg Gly Ser Tyr Met 195 200
205 Asp Phe Asn Val Glu Gly Arg Gly Val Ser Arg Pro Asp
Ile Leu Asn 210 215 220
Lys Lys Ala Glu Lys Lys Arg Leu Leu Val Ala Lys Met Ile Asp Leu 225
230 235 240 Lys His Thr Glu
Glu Trp Ala Asp Ala Ile Ala Arg Thr Asp Ser Thr 245
250 255 Val Ala Ala Gly Tyr Lys Ile Glu Pro
Tyr His His Gln Phe Trp Asn 260 265
270 Gly Asp Tyr Val Gln His Leu Arg Pro Ala Tyr Ser Phe Asn
Val Arg 275 280 285
Met Val Ser Lys Arg Thr Arg Arg Ser Glu Ser Gly Asn Lys Glu Asn 290
295 300 Leu Leu Gly Arg Tyr
Leu Ser Asp Gly Ala Thr Asn Ile Gln Leu Arg 305 310
315 320 Gly Pro Glu Tyr Tyr Asn Ile Met Pro Val
Trp Glu Trp Asp Lys Ile 325 330
335 Pro Gly Ile Thr Ser Arg Asp Tyr Leu Thr Asp Arg Pro Leu Thr
Lys 340 345 350 Leu
68401PRTArtificialSynthetic polypeptide, N(delta)80 chondroitinase B
(G[sub]106 - H[sub]506) 68Gly Asn Arg Ala Ile Gln Ala Trp Lys Ser His Gly
Pro Gly Leu Val 1 5 10
15 Ala Ile Tyr Gly Ser Tyr Asn Arg Ile Thr Ala Cys Val Phe Asp Cys
20 25 30 Phe Asp Glu
Ala Asn Ser Ala Tyr Ile Thr Thr Ser Leu Thr Glu Asp 35
40 45 Gly Lys Val Pro Gln His Cys Arg
Ile Asp His Cys Ser Phe Thr Asp 50 55
60 Lys Ile Thr Phe Asp Gln Val Ile Asn Leu Asn Asn Thr
Ala Arg Ala 65 70 75
80 Ile Lys Asp Gly Ser Val Gly Gly Pro Gly Met Tyr His Arg Val Asp
85 90 95 His Cys Phe Phe
Ser Asn Pro Gln Lys Pro Gly Asn Ala Gly Gly Gly 100
105 110 Ile Arg Ile Gly Tyr Tyr Arg Asn Asp
Ile Gly Arg Cys Leu Val Asp 115 120
125 Ser Asn Leu Phe Met Arg Gln Asp Ser Glu Ala Glu Ile Ile
Thr Ser 130 135 140
Lys Ser Gln Glu Asn Val Tyr Tyr Gly Asn Thr Tyr Leu Asn Cys Gln 145
150 155 160 Gly Thr Met Asn Phe
Arg His Gly Asp His Gln Val Ala Ile Asn Asn 165
170 175 Phe Tyr Ile Gly Asn Asp Gln Arg Phe Gly
Tyr Gly Gly Met Phe Val 180 185
190 Trp Gly Ser Arg His Val Ile Ala Cys Asn Tyr Phe Glu Leu Ser
Glu 195 200 205 Thr
Ile Lys Ser Arg Gly Asn Ala Ala Leu Tyr Leu Asn Pro Gly Ala 210
215 220 Met Ala Ser Glu His Ala
Leu Ala Phe Asp Met Leu Ile Ala Asn Asn 225 230
235 240 Ala Phe Ile Asn Val Asn Gly Tyr Ala Ile His
Phe Asn Pro Leu Asp 245 250
255 Glu Arg Arg Lys Glu Tyr Cys Ala Ala Asn Arg Leu Lys Phe Glu Thr
260 265 270 Pro His
Gln Leu Met Leu Lys Gly Asn Leu Phe Phe Lys Asp Lys Pro 275
280 285 Tyr Val Tyr Pro Phe Phe Lys
Asp Asp Tyr Phe Ile Ala Gly Lys Asn 290 295
300 Ser Trp Thr Gly Asn Val Ala Leu Gly Val Glu Lys
Gly Ile Pro Val 305 310 315
320 Asn Ile Ser Ala Asn Arg Ser Ala Tyr Lys Pro Val Lys Ile Lys Asp
325 330 335 Ile Gln Pro
Ile Glu Gly Ile Ala Leu Asp Leu Asn Ala Leu Ile Ser 340
345 350 Lys Gly Ile Thr Gly Lys Pro Leu
Ser Trp Asp Glu Val Arg Pro Tyr 355 360
365 Trp Leu Lys Glu Met Pro Gly Thr Tyr Ala Leu Thr Ala
Arg Leu Ser 370 375 380
Ala Asp Arg Ala Ala Lys Phe Lys Ala Val Ile Lys Arg Asn Lys Glu 385
390 395 400 His
69361PRTArtificialSynthetic polypeptide, N(delta)120 chondroitinase
B (I[sub]146 - H[sub]506) 69Ile Thr Thr Ser Leu Thr Glu Asp Gly Lys Val
Pro Gln His Cys Arg 1 5 10
15 Ile Asp His Cys Ser Phe Thr Asp Lys Ile Thr Phe Asp Gln Val Ile
20 25 30 Asn Leu
Asn Asn Thr Ala Arg Ala Ile Lys Asp Gly Ser Val Gly Gly 35
40 45 Pro Gly Met Tyr His Arg Val
Asp His Cys Phe Phe Ser Asn Pro Gln 50 55
60 Lys Pro Gly Asn Ala Gly Gly Gly Ile Arg Ile Gly
Tyr Tyr Arg Asn 65 70 75
80 Asp Ile Gly Arg Cys Leu Val Asp Ser Asn Leu Phe Met Arg Gln Asp
85 90 95 Ser Glu Ala
Glu Ile Ile Thr Ser Lys Ser Gln Glu Asn Val Tyr Tyr 100
105 110 Gly Asn Thr Tyr Leu Asn Cys Gln
Gly Thr Met Asn Phe Arg His Gly 115 120
125 Asp His Gln Val Ala Ile Asn Asn Phe Tyr Ile Gly Asn
Asp Gln Arg 130 135 140
Phe Gly Tyr Gly Gly Met Phe Val Trp Gly Ser Arg His Val Ile Ala 145
150 155 160 Cys Asn Tyr Phe
Glu Leu Ser Glu Thr Ile Lys Ser Arg Gly Asn Ala 165
170 175 Ala Leu Tyr Leu Asn Pro Gly Ala Met
Ala Ser Glu His Ala Leu Ala 180 185
190 Phe Asp Met Leu Ile Ala Asn Asn Ala Phe Ile Asn Val Asn
Gly Tyr 195 200 205
Ala Ile His Phe Asn Pro Leu Asp Glu Arg Arg Lys Glu Tyr Cys Ala 210
215 220 Ala Asn Arg Leu Lys
Phe Glu Thr Pro His Gln Leu Met Leu Lys Gly 225 230
235 240 Asn Leu Phe Phe Lys Asp Lys Pro Tyr Val
Tyr Pro Phe Phe Lys Asp 245 250
255 Asp Tyr Phe Ile Ala Gly Lys Asn Ser Trp Thr Gly Asn Val Ala
Leu 260 265 270 Gly
Val Glu Lys Gly Ile Pro Val Asn Ile Ser Ala Asn Arg Ser Ala 275
280 285 Tyr Lys Pro Val Lys Ile
Lys Asp Ile Gln Pro Ile Glu Gly Ile Ala 290 295
300 Leu Asp Leu Asn Ala Leu Ile Ser Lys Gly Ile
Thr Gly Lys Pro Leu 305 310 315
320 Ser Trp Asp Glu Val Arg Pro Tyr Trp Leu Lys Glu Met Pro Gly Thr
325 330 335 Tyr Ala
Leu Thr Ala Arg Leu Ser Ala Asp Arg Ala Ala Lys Phe Lys 340
345 350 Ala Val Ile Lys Arg Asn Lys
Glu His 355 360 70463PRTArtificialSynthetic
polypeptide, C(delta)19 chondroitinase B (Q[sub]26 - L[sub]488)
70Gln Val Val Ala Ser Asn Glu Thr Leu Tyr Gln Val Val Lys Glu Val 1
5 10 15 Lys Pro Gly Gly
Leu Val Gln Ile Ala Asp Gly Thr Tyr Lys Asp Val 20
25 30 Gln Leu Ile Val Ser Asn Ser Gly Lys
Ser Gly Leu Pro Ile Thr Ile 35 40
45 Lys Ala Leu Asn Pro Gly Lys Val Phe Phe Thr Gly Asp Ala
Lys Val 50 55 60
Glu Leu Arg Gly Glu His Leu Ile Leu Glu Gly Ile Trp Phe Lys Asp 65
70 75 80 Gly Asn Arg Ala Ile
Gln Ala Trp Lys Ser His Gly Pro Gly Leu Val 85
90 95 Ala Ile Tyr Gly Ser Tyr Asn Arg Ile Thr
Ala Cys Val Phe Asp Cys 100 105
110 Phe Asp Glu Ala Asn Ser Ala Tyr Ile Thr Thr Ser Leu Thr Glu
Asp 115 120 125 Gly
Lys Val Pro Gln His Cys Arg Ile Asp His Cys Ser Phe Thr Asp 130
135 140 Lys Ile Thr Phe Asp Gln
Val Ile Asn Leu Asn Asn Thr Ala Arg Ala 145 150
155 160 Ile Lys Asp Gly Ser Val Gly Gly Pro Gly Met
Tyr His Arg Val Asp 165 170
175 His Cys Phe Phe Ser Asn Pro Gln Lys Pro Gly Asn Ala Gly Gly Gly
180 185 190 Ile Arg
Ile Gly Tyr Tyr Arg Asn Asp Ile Gly Arg Cys Leu Val Asp 195
200 205 Ser Asn Leu Phe Met Arg Gln
Asp Ser Glu Ala Glu Ile Ile Thr Ser 210 215
220 Lys Ser Gln Glu Asn Val Tyr Tyr Gly Asn Thr Tyr
Leu Asn Cys Gln 225 230 235
240 Gly Thr Met Asn Phe Arg His Gly Asp His Gln Val Ala Ile Asn Asn
245 250 255 Phe Tyr Ile
Gly Asn Asp Gln Arg Phe Gly Tyr Gly Gly Met Phe Val 260
265 270 Trp Gly Ser Arg His Val Ile Ala
Cys Asn Tyr Phe Glu Leu Ser Glu 275 280
285 Thr Ile Lys Ser Arg Gly Asn Ala Ala Leu Tyr Leu Asn
Pro Gly Ala 290 295 300
Met Ala Ser Glu His Ala Leu Ala Phe Asp Met Leu Ile Ala Asn Asn 305
310 315 320 Ala Phe Ile Asn
Val Asn Gly Tyr Ala Ile His Phe Asn Pro Leu Asp 325
330 335 Glu Arg Arg Lys Glu Tyr Cys Ala Ala
Asn Arg Leu Lys Phe Glu Thr 340 345
350 Pro His Gln Leu Met Leu Lys Gly Asn Leu Phe Phe Lys Asp
Lys Pro 355 360 365
Tyr Val Tyr Pro Phe Phe Lys Asp Asp Tyr Phe Ile Ala Gly Lys Asn 370
375 380 Ser Trp Thr Gly Asn
Val Ala Leu Gly Val Glu Lys Gly Ile Pro Val 385 390
395 400 Asn Ile Ser Ala Asn Arg Ser Ala Tyr Lys
Pro Val Lys Ile Lys Asp 405 410
415 Ile Gln Pro Ile Glu Gly Ile Ala Leu Asp Leu Asn Ala Leu Ile
Ser 420 425 430 Lys
Gly Ile Thr Gly Lys Pro Leu Ser Trp Asp Glu Val Arg Pro Tyr 435
440 445 Trp Leu Lys Glu Met Pro
Gly Thr Tyr Ala Leu Thr Ala Arg Leu 450 455
460 71365PRTArtificialOTHER INFORMATION; Synthetic
polypeptide, C(delta)120 chondroitinase B (Q[sub]26 - K[sub]390)
71Gln Val Val Ala Ser Asn Glu Thr Leu Tyr Gln Val Val Lys Glu Val 1
5 10 15 Lys Pro Gly Gly
Leu Val Gln Ile Ala Asp Gly Thr Tyr Lys Asp Val 20
25 30 Gln Leu Ile Val Ser Asn Ser Gly Lys
Ser Gly Leu Pro Ile Thr Ile 35 40
45 Lys Ala Leu Asn Pro Gly Lys Val Phe Phe Thr Gly Asp Ala
Lys Val 50 55 60
Glu Leu Arg Gly Glu His Leu Ile Leu Glu Gly Ile Trp Phe Lys Asp 65
70 75 80 Gly Asn Arg Ala Ile
Gln Ala Trp Lys Ser His Gly Pro Gly Leu Val 85
90 95 Ala Ile Tyr Gly Ser Tyr Asn Arg Ile Thr
Ala Cys Val Phe Asp Cys 100 105
110 Phe Asp Glu Ala Asn Ser Ala Tyr Ile Thr Thr Ser Leu Thr Glu
Asp 115 120 125 Gly
Lys Val Pro Gln His Cys Arg Ile Asp His Cys Ser Phe Thr Asp 130
135 140 Lys Ile Thr Phe Asp Gln
Val Ile Asn Leu Asn Asn Thr Ala Arg Ala 145 150
155 160 Ile Lys Asp Gly Ser Val Gly Gly Pro Gly Met
Tyr His Arg Val Asp 165 170
175 His Cys Phe Phe Ser Asn Pro Gln Lys Pro Gly Asn Ala Gly Gly Gly
180 185 190 Ile Arg
Ile Gly Tyr Tyr Arg Asn Asp Ile Gly Arg Cys Leu Val Asp 195
200 205 Ser Asn Leu Phe Met Arg Gln
Asp Ser Glu Ala Glu Ile Ile Thr Ser 210 215
220 Lys Ser Gln Glu Asn Val Tyr Tyr Gly Asn Thr Tyr
Leu Asn Cys Gln 225 230 235
240 Gly Thr Met Asn Phe Arg His Gly Asp His Gln Val Ala Ile Asn Asn
245 250 255 Phe Tyr Ile
Gly Asn Asp Gln Arg Phe Gly Tyr Gly Gly Met Phe Val 260
265 270 Trp Gly Ser Arg His Val Ile Ala
Cys Asn Tyr Phe Glu Leu Ser Glu 275 280
285 Thr Ile Lys Ser Arg Gly Asn Ala Ala Leu Tyr Leu Asn
Pro Gly Ala 290 295 300
Met Ala Ser Glu His Ala Leu Ala Phe Asp Met Leu Ile Ala Asn Asn 305
310 315 320 Ala Phe Ile Asn
Val Asn Gly Tyr Ala Ile His Phe Asn Pro Leu Asp 325
330 335 Glu Arg Arg Lys Glu Tyr Cys Ala Ala
Asn Arg Leu Lys Phe Glu Thr 340 345
350 Pro His Gln Leu Met Leu Lys Gly Asn Leu Phe Phe Lys
355 360 365
72245PRTArtificialSynthetic polypeptide, N(delta)120 C(delta)120
chondroitinase B (I[sub]146 - K[sub]390) 72Ile Thr Thr Ser Leu Thr Glu
Asp Gly Lys Val Pro Gln His Cys Arg 1 5
10 15 Ile Asp His Cys Ser Phe Thr Asp Lys Ile Thr
Phe Asp Gln Val Ile 20 25
30 Asn Leu Asn Asn Thr Ala Arg Ala Ile Lys Asp Gly Ser Val Gly
Gly 35 40 45 Pro
Gly Met Tyr His Arg Val Asp His Cys Phe Phe Ser Asn Pro Gln 50
55 60 Lys Pro Gly Asn Ala Gly
Gly Gly Ile Arg Ile Gly Tyr Tyr Arg Asn 65 70
75 80 Asp Ile Gly Arg Cys Leu Val Asp Ser Asn Leu
Phe Met Arg Gln Asp 85 90
95 Ser Glu Ala Glu Ile Ile Thr Ser Lys Ser Gln Glu Asn Val Tyr Tyr
100 105 110 Gly Asn
Thr Tyr Leu Asn Cys Gln Gly Thr Met Asn Phe Arg His Gly 115
120 125 Asp His Gln Val Ala Ile Asn
Asn Phe Tyr Ile Gly Asn Asp Gln Arg 130 135
140 Phe Gly Tyr Gly Gly Met Phe Val Trp Gly Ser Arg
His Val Ile Ala 145 150 155
160 Cys Asn Tyr Phe Glu Leu Ser Glu Thr Ile Lys Ser Arg Gly Asn Ala
165 170 175 Ala Leu Tyr
Leu Asn Pro Gly Ala Met Ala Ser Glu His Ala Leu Ala 180
185 190 Phe Asp Met Leu Ile Ala Asn Asn
Ala Phe Ile Asn Val Asn Gly Tyr 195 200
205 Ala Ile His Phe Asn Pro Leu Asp Glu Arg Arg Lys Glu
Tyr Cys Ala 210 215 220
Ala Asn Arg Leu Lys Phe Glu Thr Pro His Gln Leu Met Leu Lys Gly 225
230 235 240 Asn Leu Phe Phe
Lys 245 732103DNAUnknownPedobacter heparinus
Chondroitinase AC nucleotide sequence 73atgaagaaat tatttgtaac
ctgtatagtc tttttctcta ttttaagtcc tgctctgctt 60attgcacagc agaccggtac
tgcagaactg attatgaagc gggtgatgct ggaccttaaa 120aagcctttgc gcaatatgga
taaggtggcg gaaaagaacc tgaatacgct gcagcctgac 180ggtagctgga aggatgtgcc
ttataaagat gatgccatga ccaattggtt gccaaacaac 240cacctgctac aattggaaac
tattatacag gcttatattg aaaaagatag tcactattat 300ggcgacgata aagtgtttga
ccagatttcc aaagctttta agtattggta tgacagcgac 360ccgaaaagcc gcaactggtg
gcacaatgaa attgccactc cgcaggccct tggtgaaatg 420ctgatcctga tgcgttacgg
taaaaagccg cttgatgaag cattggtgca taaattgacc 480gaaagaatga agcggggcga
accggagaag aaaacggggg ccaacaaaac agatatcgcc 540ctgcattact tttatcgtgc
tttgttaacg tctgatgagg ctttgctttc cttcgccgta 600aaagaattgt tttatcccgt
acagtttgta cactatgagg aaggcctgca atacgattat 660tcctacctgc agcacggtcc
gcaattacag atatcgagct acggtgccgt atttattacc 720ggggtactga aacttgccaa
ttacgttagg gatacccctt atgctttaag taccgagaaa 780ctggctatat tttcaaagta
ttaccgcgac agttatctga aagctatccg tggaagttat 840atggatttta acgtagaagg
ccgcggagta agccggccag acattctaaa taaaaaggca 900gaaaaaaaga ggttgctggt
ggcgaagatg atcgatctta agcatactga agaatgggct 960gatgcgatag ccaggacaga
tagcacagtt gcggccggct ataagattga gccctatcac 1020catcagttct ggaatggtga
ttatgtgcaa catttaagac ctgcctattc ttttaatgtt 1080cgtatggtga gtaagcggac
ccgacgcagt gaatccggca ataaagaaaa cctgctgggc 1140aggtatttat ctgatggggc
tactaacata caattgcgcg gaccagaata ctataacatt 1200atgccggtat gggaatggga
caagattcct ggcataacca gccgtgatta tttaaccgac 1260agacctttga cgaagctttg
gggagagcag gggagcaatg actttgcagg aggggtgtct 1320gatggtgtat acggggccag
tgcctacgca ttggattacg atagcttaca ggcaaagaaa 1380gcctggttct tttttgacaa
agagattgta tgtcttggtg ccggtatcaa cagcaatgcc 1440cctgaaaaca ttaccactac
ccttaaccag agctggttaa atggcccggt tataagtact 1500gcaggtaaaa ccggccgggg
taaaataaca acgtttaaag cacagggaca gttctggttg 1560ttgcacgatg cgattggtta
ttactttcct gaaggggcca accttagtct gagtacccag 1620tcgcaaaaag gcaattggtt
ccacatcaac aattcacatt caaaagatga agtttctggt 1680gatgtattta agctttggat
caaccatggt gccaggccag aaaatgcgca gtatgcttat 1740atcgttttgc cgggaataaa
caagccggaa gaaattaaaa aatataatgg aacggcaccg 1800aaagtccttg ccaataccaa
ccagctgcag gcagtttatc atcagcagtt agatatggta 1860caggctatct tctatacagc
tggaaaatta agcgtagcgg gcatagaaat tgaaacagat 1920aagccatgtg cagtgctgat
caagcacatc aatggcaagc aggtaatttg ggctgccgat 1980ccattgcaaa aagaaaagac
tgcagtgttg agcatcaggg atttaaaaac aggaaaaaca 2040aatcgggtaa aaattgattt
tccgcaacag gaatttgcag gtgcaacggt tgaactgaaa 2100tag
2103741067DNAArtificialSynthetic polynucleotide of chondroitinase AC
nucleic acid deletion N(delta)50 C(delta)275 (a[sub]220 -
t[sub]1278) 74atgccatgac caattggttg ccaaacaacc acctgctaca attggaaact
attatacagg 60cttatattga aaaagatagt cactattatg gcgacgataa agtgtttgac
cagatttcca 120aagcttttaa gtattggtat gacagcgacc cgaaaagccg caactggtgg
cacaatgaaa 180ttgccactcc gcaggccctt ggtgaaatgc tgatcctgat gcgttacggt
aaaaagccgc 240ttgatgaagc attggtgcat aaattgaccg aaagaatgaa gcggggcgaa
ccggagaaga 300aaacgggggc caacaaaaca gatatcgccc tgcattactt ttatcgtgct
ttgttaacgt 360ctgatgaggc tttgctttcc ttcgccgtaa aagaattgtt ttatcccgta
cagtttgtac 420actatgagga aggcctgcaa tacgattatt cctacctgca gcacggtccg
caattacaga 480tatcgagcta cggtgccgta tttattaccg gggtactgaa acttgccaat
tacgttaggg 540atacccctta tgctttaagt accgagaaac tggctatatt ttcaaagtat
taccgcgaca 600gttatctgaa agctatccgt ggaagttata tggattttaa cgtagaaggc
cgcggagtaa 660gccggccaga cattctaaat aaaaaggcag aaaaaaagag gttgctggtg
gcgaagatga 720tcgatcttaa gcatactgaa gaatgggctg atgcgatagc caggacagat
agcacagttg 780cggccggcta taagattgag ccctatcacc atcagttctg gaatggtgat
tatgtgcaac 840atttaagacc tgcctattct tttaatgttc gtatggtgag taagcggacc
cgacgcagtg 900aatccggcaa taaagaaaac ctgctgggca ggtatttatc tgatggggct
actaacatac 960aattgcgcgg accagaatac tataacatta tgccggtatg ggaatgggac
aagattcctg 1020gcataaccag ccgtgattat ttaaccgaca gacctttgac gaagctt
1067751521DNAUnknownPedobacter Heparinus Chondroitinase B
Chondroitinase B nucleotide sequence 75atgaagatgc tgaataaact agccggatac
ttattgccga tcatggtgct gctgaatgtg 60gcaccatgct taggtcaggt tgttgcttca
aatgaaactt tataccaggt tgtaaaggag 120gtaaaacccg gtggtctggt acagattgcc
gatgggactt ataaagatgt tcagctgatt 180gtcagcaatt caggaaaatc tggtttgccc
atcactatta aagccctgaa cccgggtaag 240gtttttttta ccggagatgc taaagtagag
ctgaggggcg agcacctgat actggaaggc 300atctggttta aagacgggaa cagagctatt
caggcatgga aatcacatgg acccggattg 360gtggctatat atggtagcta taaccgcatt
accgcatgtg tatttgattg ttttgatgaa 420gccaattctg cttacattac tacttcgctt
accgaagacg gaaaggtacc tcaacattgc 480cgcatagacc attgcagttt taccgataag
atcacttttg accaggtaat taacctgaac 540aatacagcca gagctattaa agacggttcg
gtgggaggac cggggatgta ccatcgtgtt 600gatcactgtt ttttttccaa tccgcaaaaa
ccgggtaatg ccggaggggg aatcaggatt 660ggctattacc gtaatgatat aggccgttgt
ctggtagact ctaacctgtt tatgcgtcag 720gattcggaag cagagatcat caccagcaaa
tcgcaggaaa atgtttatta tggtaatact 780tacctgaatt gccagggcac catgaacttt
cgtcacggtg atcatcaggt ggccattaac 840aatttttata taggcaatga ccagcgattt
ggatacgggg gaatgtttgt ttggggaagc 900aggcatgtca tagcctgtaa ttattttgag
ctgtccgaaa ccataaagtc gagggggaac 960gccgcattgt atttaaaccc cggtgctatg
gcttcggagc atgctcttgc tttcgatatg 1020ttgatagcca acaacgcttt catcaatgta
aatgggtatg ccatccattt taatccattg 1080gatgagcgca gaaaagaata ttgtgcagcc
aataggctta agttcgaaac cccgcaccag 1140ctaatgttaa aaggcaatct tttctttaag
gataaacctt atgtttaccc attttttaaa 1200gatgattatt ttatagcagg gaaaaatagc
tggactggta atgtagcctt aggtgtggaa 1260aagggaatcc ctgttaacat ttcggccaat
aggtctgcct ataagccggt aaaaattaaa 1320gatatccagc ccatagaagg aatcgctctt
gatctcaatg cgctgatcag caaaggcatt 1380acaggaaagc cccttagctg ggatgaagta
aggccctact ggttaaaaga aatgcccggg 1440acgtatgctt taacggccag gctttctgca
gatagggctg caaagtttaa agccgtaatt 1500aaaagaaata aagagcactg a
152176735DNAArtificialSynthetic
polynucleotide of chondroitinase B nucleic acid deletion N(delta)120
C(delta)120 (a[sub]436 - g[sub]1170) 76attactactt cgcttaccga
agacggaaag gtacctcaac attgccgcat agaccattgc 60agttttaccg ataagatcac
ttttgaccag gtaattaacc tgaacaatac agccagagct 120attaaagacg gttcggtggg
aggaccgggg atgtaccatc gtgttgatca ctgttttttt 180tccaatccgc aaaaaccggg
taatgccgga gggggaatca ggattggcta ttaccgtaat 240gatataggcc gttgtctggt
agactctaac ctgtttatgc gtcaggattc ggaagcagag 300atcatcacca gcaaatcgca
ggaaaatgtt tattatggta atacttacct gaattgccag 360ggcaccatga actttcgtca
cggtgatcat caggtggcca ttaacaattt ttatataggc 420aatgaccagc gatttggata
cgggggaatg tttgtttggg gaagcaggca tgtcatagcc 480tgtaattatt ttgagctgtc
cgaaaccata aagtcgaggg ggaacgccgc attgtattta 540aaccccggtg ctatggcttc
ggagcatgct cttgctttcg atatgttgat agccaacaac 600gctttcatca atgtaaatgg
gtatgccatc cattttaatc cattggatga gcgcagaaaa 660gaatattgtg cagccaatag
gcttaagttc gaaaccccgc accagctaat gttaaaaggc 720aatcttttct ttaag
735773980DNAArtificialSynthetic polynucleotide Chondroitinase ABC I
LOCUS (I29953) 77ggaattccat cactcaatca ttaaatttag gcacaacgat gggctatcag
cgttatgaca 60aatttaatga aggacgcatt ggtttcactg ttagccagcg tttctaagga
gaaaaataat 120gccgatattt cgttttactg cacttgcaat gacattgggg ctattatcag
cgccttataa 180cgcgatggca gccaccagca atcctgcatt tgatcctaaa aatctgatgc
agtcagaaat 240ttaccatttt gcacaaaata acccattagc agacttctca tcagataaaa
actcaatact 300aacgttatct gataaacgta gcattatggg aaaccaatct cttttatgga
aatggaaagg 360tggtagtagc tttactttac ataaaaaact gattgtcccc accgataaag
aagcatctaa 420agcatgggga cgctcatcta cccccgtttt ctcattttgg ctttacaatg
aaaaaccgat 480tgatggttat cttactatcg atttcggaga aaaactcatt tcaaccagtg
aggctcaggc 540aggctttaaa gtaaaattag atttcactgg ctggcgtgct gtgggagtct
ctttaaataa 600cgatcttgaa aatcgagaga tgaccttaaa tgcaaccaat acctcctctg
atggtactca 660agacagcatt gggcgttctt taggtgctaa agtcgatagt attcgtttta
aagcgccttc 720taatgtgagt cagggtgaaa tctatatcga ccgtattatg ttttctgtcg
atgatgctcg 780ctaccaatgg tctgattatc aagtaaaaac tcgcttatca gaacctgaaa
ttcaatttca 840caacgtaaag ccacaactac ctgtaacacc tgaaaattta gcggccattg
atcttattcg 900ccaacgtcta attaatgaat ttgtcggagg tgaaaaagag acaaacctcg
cattagaaga 960gaatatcagc aaattaaaaa gtgatttcga tgctcttaat attcacactt
tagcaaatgg 1020tggaacgcaa ggcagacatc tgatcactga taaacaaatc attatttatc
aaccagagaa 1080tcttaactcc caagataaac aactatttga taattatgtt attttaggta
attacacgac 1140attaatgttt aatattagcc gtgcttatgt gctggaaaaa gatcccacac
aaaaggcgca 1200actaaagcag atgtacttat taatgacaaa gcatttatta gatcaaggct
ttgttaaagg 1260gagtgcttta gtgacaaccc atcactgggg atacagttct cgttggtggt
atatttccac 1320gttattaatg tctgatgcac taaaagaagc gaacctacaa actcaagttt
atgattcatt 1380actgtggtat tcacgtgagt ttaaaagtag ttttgatatg aaagtaagtg
ctgatagctc 1440tgatctagat tatttcaata ccttatctcg ccaacattta gccttattat
tactagagcc 1500tgatgatcaa aagcgtatca acttagttaa tactttcagc cattatatca
ctggcgcatt 1560aacgcaagtg ccaccgggtg gtaaagatgg tttacgccct gatggtacag
catggcgaca 1620tgaaggcaac tatccgggct actctttccc agcctttaaa aatgcctctc
agcttattta 1680tttattacgc gatacaccat tttcagtggg tgaaagtggt tggaataacc
tgaaaaaagc 1740gatggtttca gcgtggatct acagtaatcc agaagttgga ttaccgcttg
caggaagaca 1800cccttttaac tcaccttcgt taaaatcagt cgctcaaggc tattactggc
ttgccatgtc 1860tgcaaaatca tcgcctgata aaacacttgc atctatttat cttgcgatta
gtgataaaac 1920acaaaatgaa tcaactgcta tttttggaga aactattaca ccagcgtctt
tacctcaagg 1980tttctatgcc tttaatggcg gtgcttttgg tattcatcgt tggcaagata
aaatggtgac 2040actgaaagct tataacacca atgtttggtc atctgaaatt tataacaaag
ataaccgtta 2100tggccgttac caaagtcatg gtgtcgctca aatagtgagt aatggctcgc
agctttcaca 2160gggctatcag caagaaggtt gggattggaa tagaatgcaa ggggcaacca
ctattcacct 2220tcctcttaaa gacttagaca gtcctaaacc tcatacctta atgcaacgtg
gagagcgtgg 2280atttagcgga acatcatccc ttgaaggtca atatggcatg atggcattcg
atcttattta 2340tcccgccaat cttgagcgtt ttgatcctaa tttcactgcg aaaaagagtg
tattagccgc 2400tgataatcac ttaattttta ttggtagcaa tataaatagt agtgataaaa
ataaaaatgt 2460tgaaacgacc ttattccaac atgccattac tccaacatta aatacccttt
ggattaatgg 2520acaaaagata gaaaacatgc cttatcaaac aacacttcaa caaggtgatt
ggttaattga 2580tagcaatggc aatggttact taattactca agcagaaaaa gtaaatgtaa
gtcgccaaca 2640tcaggtttca gcggaaaata aaaatcgcca accgacagaa ggaaacttta
gctcggcatg 2700gatcgatcac agcactcgcc ccaaagatgc cagttatgag tatatggtct
ttttagatgc 2760gacacctgaa aaaatgggag agatggcaca aaaattccgt gaaaataatg
ggttatatca 2820ggttcttcgt aaggataaag acgttcatat tattctcgat aaactcagca
atgtaacggg 2880atatgccttt tatcagccag catcaattga agacaaatgg atcaaaaagg
ttaataaacc 2940tgcaattgtg atgactcatc gacaaaaaga cactcttatt gtcagtgcag
ttacacctga 3000tttaaatatg actcgccaaa aagcagcaac tcctgtcacc atcaatgtca
cgattaatgg 3060caaatggcaa tctgctgata aaaatagtga agtgaaatat caggtttctg
gtgataacac 3120tgaactgacg tttacgagtt actttggtat tccacaagaa atcaaactct
cgccactccc 3180ttgatttaat caaaagaacg ctcttgcgtt ccttttttat ttgcaggaaa
tctgattatg 3240ctaataaaaa accctttagc ccacgcggtt acattaagcc tctgtttatc
attacccgca 3300caagcattac ccactctgtc tcatgaagct ttcggcgata tttatctttt
tgaaggtgaa 3360ttacccaata cccttaccac ttcaaataat aatcaattat cgctaagcaa
acagcatgct 3420aaagatggtg aacaatcact caaatggcaa tatcaaccac aagcaacatt
aacactaaat 3480aatattgtta attaccaaga tgataaaaat acagccacac cactcacttt
tatgatgtgg 3540atttataatg aaaaacctca atcttcccca ttaacgttag catttaaaca
aaataataaa 3600attgcactaa gttttaatgc tgaacttaat tttacggggt ggcgaggtat
tgctgttcct 3660tttcgtgata tgcaaggctc tgcgacaggt caacttgatc aattagtgat
caccgctcca 3720aaccaagccg gaacactctt ttttgatcaa atcatcatga gtgtaccgtt
agacaatcgt 3780tgggcagtac ctgactatca aacaccttac gtaaataacg cagtaaacac
gatggttagt 3840aaaaactgga gtgcattatt gatgtacgat cagatgtttc aagcccatta
ccctacttta 3900aacttcgata ctgaatttcg cgatgaccaa acagaaatgg cttcgattta
tcagcgcttt 3960gaatattatc aaggaattcc
39807815PRTArtificialOTHER INFORMATION; Synthetic polypeptide,
HIV TAT Sequence and Gly penta linker 78Gly Arg Lys Lys Arg Arg Gln
Arg Arg Arg Gly Gly Gly Gly Gly 1 5 10
15 7945DNAArtificialSynthetic polynucleotide, HIV-1 Tat
Sequence and Gly penta linker nucleic acid 79ggtcgtaaaa agcgtcgtca
acgtcgtcgt ggtggtggtg gtggt
45802973DNAArtificialSynthetic polynucleotide, Chondroitinase ABC II
nucleic acid 80ttacccactc tgtctcatga agctttcggc gatatttatc tttttgaagg
cgaattaccc 60aatatcctta ccacttcaaa taataatcaa ttatcgctaa gcaaacagca
tgctaaagat 120ggtgaacaat cactcaaatg gcaatatcaa ccacaagcaa cattaacact
aaataatatt 180gttaattacc aagatgataa aaatacagcc acaccactca cttttatgat
gtggatttat 240aatgaaaaac ctcaatcttc cccattaacg ttagcattta aacaaaataa
taaaattgca 300ctaagtttta atgctgaact taattttacg gggtggcgag gtattgctgt
tccttttcgt 360gatatgcaag gctctgcgac aggtcaactt gatcaattag tgatcaccgc
tccaaaccaa 420gccggaacac tcttttttga tcaaatcatc atgagtgtac cgttagacaa
tcgttgggca 480gtacctgact atcaaacacc ttacgtaaat aacgcagtaa acacgatggt
tagtaaaaac 540tggagtgcat tattgatgta cgatcagatg tttcaagccc attaccctac
tttaaacttc 600gatactgaat ttcgcgatga ccaaacagaa atggcttcga tttatcagcg
ctttgaatat 660tatcaaggaa ttcgtagtga taaaaaaatt actccagata tgctagataa
acatttagcg 720ttatgggaaa aattggggtt aacacaacac gctgatggct caatcacagg
aaaagccctt 780gatcacccta accggcaaca ttttatgaaa gtcgaaggtg tatttagtga
ggggactcaa 840aaagcattac ttgatgccaa tatgctaaga gatgtgggca aaacgcttct
tcaaactgct 900atttacttgc gtagcgattc attatcagca actggtagaa aaaaattaga
agagcgctat 960ttattaggta ctcgttatgt ccttgaacaa ggttttacac gaggaagtgg
ttatcaaatt 1020attactcatg ttggttacca aaccagagaa ctttttgatg catggtttat
tggccgtcat 1080gttcttgcaa aaaataacct tttagccccc actcaacaag ctatgatgtg
gtacaacgcc 1140acaggacgta tttttgaaaa agataatgaa attgttgatg caaatgtcga
tattctcaat 1200actcaattgc aatggatgat aaaaagctta ttgatgctac cggattatca
acaacgtcaa 1260caagccttag cgcaactgca aagttggcta aataaaacca ttctaagctc
aaaaggtgtt 1320gctggcggtt tcaaatctga tggttctatt tttcaccatt cacaacatta
ccccgcttat 1380gctaaagatg catttggtgg tttagcaccc agtgtttatg cattaagtga
ttcacctttt 1440cgcttatcta cttcagcaca tgagcattta aaagatgttt tgttaaaaat
gcggatctac 1500accaaagaga cacaaattcc tgtggtatta agtggtcgtc atccaactgg
gttgcataaa 1560atagggatcg cgccatttaa atggatggca ttagcaggaa ccccagatgg
caaacaaaag 1620ttagatacca cattatccgc cgcttatgca aacttagaca acaaaacgca
ttttgaaggc 1680attaacgctg aaagtgagcc agtcggcgca tgggcaatga attatgcatc
aatggcaata 1740caacgaagag catcgaccca atcaccacaa caaagctggc tcgccatagc
gcgcggtttt 1800agccgttatc ttgttggtaa tgaaagctat gaaaataaca accgttatgg
tcgttattta 1860caatatggac aattggaaat tattccagct gatttaactc aatcagggtt
tagccatgct 1920ggatgggatt ggaatagata tccaggtaca acaactattc atcttcccta
taacgaactt 1980gaagcaaaac ttaatcaatt acctgctgca ggtattgaag aaatgttgct
ttcaacagaa 2040agttactctg gtgcaaatac ccttaataat aacagtatgt ttgccatgaa
attacacggt 2100cacagtaaat atcaacaaca aagcttaagg gcaaataaat cctatttctt
atttgataat 2160agagttattg ctttaggctc aggtattgaa aatgatgata aacaacatac
gaccgaaaca 2220acactattcc agtttgccgt ccctaaatta cagtcagtga tcattaatgg
caaaaaggta 2280aatcaattag atactcaatt aactttaaat aatgcagata cattaattga
tcctgccggc 2340aatttatata agctcactaa aggacaaact gtaaaattta gttatcaaaa
acaacattca 2400cttgatgata gaaattcaaa accaacagaa caattatttg caacagctgt
tatttctcat 2460ggtaaggcac cgagtaatga aaattatgaa tatgcaatag ctatcgaagc
acaaaataat 2520aaagctccca aatacacagt attacaacat aatgatcagc tccatgcggt
aaaagataaa 2580ataacccaag aagagggata tggttttttt gaagccacta agttaaaatc
agcggatgca 2640acattattat ccagtgatgc gccggttatg gtcatggcta aaatacaaaa
tcagcaatta 2700acattaagta ttgttaatcc tgatttaaat ttatatcaag gtagagaaaa
agatcaattt 2760gatgataaag gtaatcaaat cgaagttagt gtttattctc gtcattggct
tacagcagaa 2820tcgcaatcaa caaatagtac tattaccgta aaaggaatat ggaaattaac
gacacctcaa 2880cccggtgtta ttattaagca ccacaataac aacactctta ttacgacaac
aaccatacag 2940gcaacaccta ctgttattaa tttagttaag taa
297381977PRTArtificialSynthetic polypeptide, N(delta)20
chondroitinase ABCI having gwra and dalni sequences 81Ala Gln Asn Asn Pro
Leu Ala Asp Phe Ser Ser Asp Lys Asn Ser Ile 1 5
10 15 Leu Thr Leu Ser Asp Lys Arg Ser Ile Met
Gly Asn Gln Ser Leu Leu 20 25
30 Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His Lys Lys Leu
Ile 35 40 45 Val
Pro Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly Arg Ser Ser Thr 50
55 60 Pro Val Phe Ser Phe Trp
Leu Tyr Asn Glu Lys Pro Ile Asp Gly Tyr 65 70
75 80 Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile Ser
Thr Ser Glu Ala Gln 85 90
95 Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp Arg Ala Val Gly
100 105 110 Val Ser
Leu Asn Asn Asp Leu Glu Asn Arg Glu Met Thr Leu Asn Ala 115
120 125 Thr Asn Thr Ser Ser Asp Gly
Thr Gln Asp Ser Ile Gly Arg Ser Leu 130 135
140 Gly Ala Lys Val Asp Ser Ile Arg Phe Lys Ala Pro
Ser Asn Val Ser 145 150 155
160 Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala
165 170 175 Arg Tyr Gln
Trp Ser Asp Tyr Gln Val Lys Thr Arg Leu Ser Glu Pro 180
185 190 Glu Ile Gln Phe His Asn Val Lys
Pro Gln Leu Pro Val Thr Pro Glu 195 200
205 Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu Ile
Asn Glu Phe 210 215 220
Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu Glu Asn Ile Ser 225
230 235 240 Lys Leu Lys Ser
Asp Phe Asp Ala Leu Asn Ile His Thr Leu Ala Asn 245
250 255 Gly Gly Thr Gln Gly Arg His Leu Ile
Thr Asp Lys Gln Ile Ile Ile 260 265
270 Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe
Asp Asn 275 280 285
Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe Asn Ile Ser Arg 290
295 300 Ala Tyr Val Leu Glu
Lys Asp Pro Thr Gln Lys Ala Gln Leu Lys Gln 305 310
315 320 Met Tyr Leu Leu Met Thr Lys His Leu Leu
Asp Gln Gly Phe Val Lys 325 330
335 Gly Ser Ala Leu Val Thr Thr His His Trp Gly Tyr Ser Ser Arg
Trp 340 345 350 Trp
Tyr Ile Ser Thr Leu Leu Met Ser Asp Ala Leu Lys Glu Ala Asn 355
360 365 Leu Gln Thr Gln Val Tyr
Asp Ser Leu Leu Trp Tyr Ser Arg Glu Phe 370 375
380 Lys Ser Ser Phe Asp Met Lys Val Ser Ala Asp
Ser Ser Asp Leu Asp 385 390 395
400 Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu
405 410 415 Pro Asp
Asp Gln Lys Arg Ile Asn Leu Val Asn Thr Phe Ser His Tyr 420
425 430 Ile Thr Gly Ala Leu Thr Gln
Val Pro Pro Gly Gly Lys Asp Gly Leu 435 440
445 Arg Pro Asp Gly Thr Ala Trp Arg His Glu Gly Asn
Tyr Pro Gly Tyr 450 455 460
Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg 465
470 475 480 Asp Thr Pro
Phe Ser Val Gly Glu Ser Gly Trp Asn Asn Leu Lys Lys 485
490 495 Ala Met Val Ser Ala Trp Ile Tyr
Ser Asn Pro Glu Val Gly Leu Pro 500 505
510 Leu Ala Gly Arg His Pro Phe Asn Ser Pro Ser Leu Lys
Ser Val Ala 515 520 525
Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser Ser Pro Asp Lys 530
535 540 Thr Leu Ala Ser
Ile Tyr Leu Ala Ile Ser Asp Lys Thr Gln Asn Glu 545 550
555 560 Ser Thr Ala Ile Phe Gly Glu Thr Ile
Thr Pro Ala Ser Leu Pro Gln 565 570
575 Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile His Arg
Trp Gln 580 585 590
Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn Val Trp Ser Ser
595 600 605 Glu Ile Tyr Asn
Lys Asp Asn Arg Tyr Gly Arg Tyr Gln Ser His Gly 610
615 620 Val Ala Gln Ile Val Ser Asn Gly
Ser Gln Leu Ser Gln Gly Tyr Gln 625 630
635 640 Gln Glu Gly Trp Asp Trp Asn Arg Met Glu Gly Ala
Thr Thr Ile His 645 650
655 Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His Thr Leu Met Gln
660 665 670 Arg Gly Glu
Arg Gly Phe Ser Gly Thr Ser Ser Leu Glu Gly Gln Tyr 675
680 685 Gly Met Met Ala Phe Asn Leu Ile
Tyr Pro Ala Asn Leu Glu Arg Phe 690 695
700 Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala Ala
Asp Asn His 705 710 715
720 Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp Lys Asn Lys Asn
725 730 735 Val Glu Thr Thr
Leu Phe Gln His Ala Ile Thr Pro Thr Leu Asn Thr 740
745 750 Leu Trp Ile Asn Gly Gln Lys Ile Glu
Asn Met Pro Tyr Gln Thr Thr 755 760
765 Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly
Tyr Leu 770 775 780
Ile Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln His Gln Val Ser 785
790 795 800 Ala Glu Asn Lys Asn
Arg Gln Pro Thr Glu Gly Asn Phe Ser Ser Ala 805
810 815 Trp Ile Asp His Ser Thr Arg Pro Lys Asp
Ala Ser Tyr Glu Tyr Met 820 825
830 Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln
Lys 835 840 845 Phe
Arg Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg Lys Asp Lys Asp 850
855 860 Val His Ile Ile Leu Asp
Lys Leu Ser Asn Val Thr Gly Tyr Ala Phe 865 870
875 880 Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp Ile
Lys Lys Val Asn Lys 885 890
895 Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr Leu Ile Val Ser
900 905 910 Ala Val
Thr Pro Asp Leu Asn Met Thr Arg Gln Lys Ala Ala Thr Pro 915
920 925 Val Thr Ile Asn Val Thr Ile
Asn Gly Lys Trp Gln Ser Ala Asp Lys 930 935
940 Asn Ser Glu Val Lys Tyr Gln Val Ser Gly Asp Asn
Thr Glu Leu Thr 945 950 955
960 Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys Leu Ser Pro Leu
965 970 975 Pro
82937PRTArtificialSynthetic polypeptide, N(delta)60 chondroitinase
ABCI having gwra and dalni sequences 82Phe Thr Leu His Lys Lys Leu Ile
Val Pro Thr Asp Lys Glu Ala Ser 1 5 10
15 Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser Phe
Trp Leu Tyr 20 25 30
Asn Glu Lys Pro Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys
35 40 45 Leu Ile Ser Thr
Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp 50
55 60 Phe Thr Gly Trp Arg Ala Val Gly
Val Ser Leu Asn Asn Asp Leu Glu 65 70
75 80 Asn Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser
Ser Asp Gly Thr 85 90
95 Gln Asp Ser Ile Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg
100 105 110 Phe Lys Ala
Pro Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp Arg 115
120 125 Ile Met Phe Ser Val Asp Asp Ala
Arg Tyr Gln Trp Ser Asp Tyr Gln 130 135
140 Val Lys Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe His
Asn Val Lys 145 150 155
160 Pro Gln Leu Pro Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile
165 170 175 Arg Gln Arg Leu
Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn 180
185 190 Leu Ala Leu Glu Glu Asn Ile Ser Lys
Leu Lys Ser Asp Phe Asp Ala 195 200
205 Leu Asn Ile His Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg
His Leu 210 215 220
Ile Thr Asp Lys Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser 225
230 235 240 Gln Asp Lys Gln Leu
Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr 245
250 255 Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr
Val Leu Glu Lys Asp Pro 260 265
270 Thr Gln Lys Ala Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys
His 275 280 285 Leu
Leu Asp Gln Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His 290
295 300 His Trp Gly Tyr Ser Ser
Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met 305 310
315 320 Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr
Gln Val Tyr Asp Ser 325 330
335 Leu Leu Trp Tyr Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val
340 345 350 Ser Ala
Asp Ser Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln 355
360 365 His Leu Ala Leu Leu Leu Leu
Glu Pro Asp Asp Gln Lys Arg Ile Asn 370 375
380 Leu Val Asn Thr Phe Ser His Tyr Ile Thr Gly Ala
Leu Thr Gln Val 385 390 395
400 Pro Pro Gly Gly Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg
405 410 415 His Glu Gly
Asn Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn Ala 420
425 430 Ser Gln Leu Ile Tyr Leu Leu Arg
Asp Thr Pro Phe Ser Val Gly Glu 435 440
445 Ser Gly Trp Asn Asn Leu Lys Lys Ala Met Val Ser Ala
Trp Ile Tyr 450 455 460
Ser Asn Pro Glu Val Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn 465
470 475 480 Ser Pro Ser Leu
Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met 485
490 495 Ser Ala Lys Ser Ser Pro Asp Lys Thr
Leu Ala Ser Ile Tyr Leu Ala 500 505
510 Ile Ser Asp Lys Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly
Glu Thr 515 520 525
Ile Thr Pro Ala Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly 530
535 540 Ala Phe Gly Ile His
Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala 545 550
555 560 Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile
Tyr Asn Lys Asp Asn Arg 565 570
575 Tyr Gly Arg Tyr Gln Ser His Gly Val Ala Gln Ile Val Ser Asn
Gly 580 585 590 Ser
Gln Leu Ser Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg 595
600 605 Met Glu Gly Ala Thr Thr
Ile His Leu Pro Leu Lys Asp Leu Asp Ser 610 615
620 Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu
Arg Gly Phe Ser Gly 625 630 635
640 Thr Ser Ser Leu Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile
645 650 655 Tyr Pro
Ala Asn Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys 660
665 670 Ser Val Leu Ala Ala Asp Asn
His Leu Ile Phe Ile Gly Ser Asn Ile 675 680
685 Asn Ser Ser Asp Lys Asn Lys Asn Val Glu Thr Thr
Leu Phe Gln His 690 695 700
Ala Ile Thr Pro Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile 705
710 715 720 Glu Asn Met
Pro Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile 725
730 735 Asp Ser Asn Gly Asn Gly Tyr Leu
Ile Thr Gln Ala Glu Lys Val Asn 740 745
750 Val Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys Asn
Arg Gln Pro 755 760 765
Thr Glu Gly Asn Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro 770
775 780 Lys Asp Ala Ser
Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu 785 790
795 800 Lys Met Gly Glu Met Ala Gln Lys Phe
Arg Glu Asn Asn Gly Leu Tyr 805 810
815 Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile Leu Asp
Lys Leu 820 825 830
Ser Asn Val Thr Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp
835 840 845 Lys Trp Ile Lys
Lys Val Asn Lys Pro Ala Ile Val Met Thr His Arg 850
855 860 Gln Lys Asp Thr Leu Ile Val Ser
Ala Val Thr Pro Asp Leu Asn Met 865 870
875 880 Thr Arg Gln Lys Ala Ala Thr Pro Val Thr Ile Asn
Val Thr Ile Asn 885 890
895 Gly Lys Trp Gln Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val
900 905 910 Ser Gly Asp
Asn Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro 915
920 925 Gln Glu Ile Lys Leu Ser Pro Leu
Pro 930 935 83858PRTArtificialSynthetic
polypeptide, N(delta)60 C(delta)80 chondroitinase ABCI having gwra
and dalni sequences 83Phe Thr Leu His Lys Lys Leu Ile Val Pro Thr Asp Lys
Glu Ala Ser 1 5 10 15
Lys Ala Trp Gly Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr
20 25 30 Asn Glu Lys Pro
Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys 35
40 45 Leu Ile Ser Thr Ser Glu Ala Gln Ala
Gly Phe Lys Val Lys Leu Asp 50 55
60 Phe Thr Gly Trp Arg Ala Val Gly Val Ser Leu Asn Asn
Asp Leu Glu 65 70 75
80 Asn Arg Glu Met Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr
85 90 95 Gln Asp Ser Ile
Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg 100
105 110 Phe Lys Ala Pro Ser Asn Val Ser Gln
Gly Glu Ile Tyr Ile Asp Arg 115 120
125 Ile Met Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp
Tyr Gln 130 135 140
Val Lys Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys 145
150 155 160 Pro Gln Leu Pro Val
Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile 165
170 175 Arg Gln Arg Leu Ile Asn Glu Phe Val Gly
Gly Glu Lys Glu Thr Asn 180 185
190 Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp
Ala 195 200 205 Leu
Asn Ile His Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu 210
215 220 Ile Thr Asp Lys Gln Ile
Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser 225 230
235 240 Gln Asp Lys Gln Leu Phe Asp Asn Tyr Val Ile
Leu Gly Asn Tyr Thr 245 250
255 Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro
260 265 270 Thr Gln
Lys Ala Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His 275
280 285 Leu Leu Asp Gln Gly Phe Val
Lys Gly Ser Ala Leu Val Thr Thr His 290 295
300 His Trp Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser
Thr Leu Leu Met 305 310 315
320 Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser
325 330 335 Leu Leu Trp
Tyr Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val 340
345 350 Ser Ala Asp Ser Ser Asp Leu Asp
Tyr Phe Asn Thr Leu Ser Arg Gln 355 360
365 His Leu Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln Lys
Arg Ile Asn 370 375 380
Leu Val Asn Thr Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val 385
390 395 400 Pro Pro Gly Gly
Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg 405
410 415 His Glu Gly Asn Tyr Pro Gly Tyr Ser
Phe Pro Ala Phe Lys Asn Ala 420 425
430 Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val
Gly Glu 435 440 445
Ser Gly Trp Asn Asn Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr 450
455 460 Ser Asn Pro Glu Val
Gly Leu Pro Leu Ala Gly Arg His Pro Phe Asn 465 470
475 480 Ser Pro Ser Leu Lys Ser Val Ala Gln Gly
Tyr Tyr Trp Leu Ala Met 485 490
495 Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu
Ala 500 505 510 Ile
Ser Asp Lys Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr 515
520 525 Ile Thr Pro Ala Ser Leu
Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly 530 535
540 Ala Phe Gly Ile His Arg Trp Gln Asp Lys Met
Val Thr Leu Lys Ala 545 550 555
560 Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg
565 570 575 Tyr Gly
Arg Tyr Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly 580
585 590 Ser Gln Leu Ser Gln Gly Tyr
Gln Gln Glu Gly Trp Asp Trp Asn Arg 595 600
605 Met Glu Gly Ala Thr Thr Ile His Leu Pro Leu Lys
Asp Leu Asp Ser 610 615 620
Pro Lys Pro His Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly 625
630 635 640 Thr Ser Ser
Leu Glu Gly Gln Tyr Gly Met Met Ala Phe Asn Leu Ile 645
650 655 Tyr Pro Ala Asn Leu Glu Arg Phe
Asp Pro Asn Phe Thr Ala Lys Lys 660 665
670 Ser Val Leu Ala Ala Asp Asn His Leu Ile Phe Ile Gly
Ser Asn Ile 675 680 685
Asn Ser Ser Asp Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His 690
695 700 Ala Ile Thr Pro
Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile 705 710
715 720 Glu Asn Met Pro Tyr Gln Thr Thr Leu
Gln Gln Gly Asp Trp Leu Ile 725 730
735 Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys
Val Asn 740 745 750
Val Ser Arg Gln His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro
755 760 765 Thr Glu Gly Asn
Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro 770
775 780 Lys Asp Ala Ser Tyr Glu Tyr Met
Val Phe Leu Asp Ala Thr Pro Glu 785 790
795 800 Lys Met Gly Glu Met Ala Gln Lys Phe Arg Glu Asn
Asn Gly Leu Tyr 805 810
815 Gln Val Leu Arg Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu
820 825 830 Ser Asn Val
Thr Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp 835
840 845 Lys Trp Ile Lys Lys Val Asn Lys
Pro Ala 850 855
842976DNAArtificialSynthetic polynucleotide, TAT-Chondroitinase ABC
I N(delta)20 Nucleic Acid 84ggtcgtaaaa agcgtcgtca acgtcgtcgt cctcctcaat
gcgcacaaaa taacccatta 60gcagacttct catcagataa aaactcaata ctaacgttat
ctgataaacg tagcattatg 120ggaaaccaat ctcttttatg gaaatggaaa ggtggtagta
gctttacttt acataaaaaa 180ctgattgtcc ccaccgataa agaagcatct aaagcatggg
gacgctcatc cacccccgtt 240ttctcatttt ggctttacaa tgaaaaaccg attgatggtt
atcttactat cgatttcgga 300gaaaaactca tttcaaccag tgaggctcag gcaggcttta
aagtaaaatt agatttcact 360ggctggcgta ctgtgggagt ctctttaaat aacgatcttg
aaaatcgaga gatgacctta 420aatgcaacca atacctcctc tgatggtact caagacagca
ttgggcgttc tttaggtgct 480aaagtcgata gtattcgttt taaagcgcct tctaatgtga
gtcagggtga aatctatatc 540gaccgtatta tgttttctgt cgatgatgct cgctaccaat
ggtctgatta tcaagtaaaa 600actcgcttat cagaacctga aattcaattt cacaacgtaa
agccacaact acctgtaaca 660cctgaaaatt tagcggccat tgatcttatt cgccaacgtc
taattaatga atttgtcgga 720ggtgaaaaag agacaaacct cgcattagaa gagaatatca
gcaaattaaa aagtgatttc 780gatgctctta atactcacac tttagcaaat ggtggaacgc
aaggcagaca tctgatcact 840gataaacaaa tcattattta tcaaccagag aatcttaact
ctcaagataa acaactattt 900gataattatg ttattttagg taattacacg acattaatgt
ttaatattag ccgtgcttat 960gtgctggaaa aagatcccac acaaaaggcg caactaaagc
agatgtactt attaatgaca 1020aagcatttat tagatcaagg ctttgttaaa gggagtgctt
tagtgacaac ccatcactgg 1080ggatacagtt ctcgttggtg gtatatttcc acgttattaa
tgtctgatgc actaaaagaa 1140gcgaacctac aaactcaagt ttatgattca ttactgtggt
attcacgtga gtttaaaagt 1200agttttgata tgaaagtaag tgctgatagc tctgatctag
attatttcaa taccttatct 1260cgccaacatt tagccttatt actactagag cctgatgatc
aaaagcgtat caacttagtt 1320aatactttca gccattatat cactggcgca ttaacgcaag
tgccaccggg tggtaaagat 1380ggtttacgcc ctgatggtac agcatggcga catgaaggca
actatccggg ctactctttc 1440ccagccttta aaaatgcctc tcagcttatt tatttattac
gcgatacacc attttcagtg 1500ggtgaaagtg gttggaatag cctgaaaaaa gcgatggttt
cagcgtggat ctacagtaat 1560ccagaagttg gattaccgct tgcaggaaga caccctctta
actcaccttc gttaaaatca 1620gtcgctcaag gctattactg gcttgccatg tctgcaaaat
catcgcctga taaaacactt 1680gcatctattt atcttgcgat tagtgataaa acacaaaatg
aatcaactgc tatttttgga 1740gaaactatta caccagcgtc tttacctcaa ggtttctatg
cctttaatgg cggtgctttt 1800ggtattcatc gttggcaaga taaaatggtg acactgaaag
cttataacac caatgtttgg 1860tcatctgaaa tttataacaa agataaccgt tatggccgtt
accaaagtca tggtgtcgct 1920caaatagtga gtaatggctc gcagctttca cagggctatc
agcaagaagg ttgggattgg 1980aatagaatgc caggggcaac cactatccac cttcctctta
aagacttaga cagtcctaaa 2040cctcatacct taatgcaacg tggagagcgt ggatttagcg
gaacatcatc ccttgaaggt 2100caatatggca tgatggcatt cgatcttatt tatcccgcca
atcttgagcg ttttgatcct 2160aatttcactg cgaaaaagag tgtattagcc gctgataatc
acttaatttt tattggtagc 2220aatataaata gtagtgataa aaataaaaat gttgaaacga
ccttattcca acatgccatt 2280actccaacat taaataccct ttggattaat ggacaaaaga
tagaaaacat gccttatcaa 2340acaacacttc aacaaggtga ttggttaatt gatagcaatg
gcaatggtta cttaattact 2400caagcagaaa aagtaaatgt aagtcgccaa catcaggttt
cagcggaaaa taaaaatcgc 2460caaccgacag aaggaaactt tagctcggca tggatcgatc
acagcactcg ccccaaagat 2520gccagttatg agtatatggt ctttttagat gcgacacctg
aaaaaatggg agagatggca 2580caaaaattcc gtgaaaataa tgggttatat caggttcttc
gtaaggataa agacgttcat 2640attattctcg ataaactcag caatgtaacg ggatatgcct
tttatcagcc agcatcaatt 2700gaagacaaat ggatcaaaaa ggttaataaa cctgcaattg
tgatgactca tcgacaaaaa 2760gacactctta ttgtcagtgc agttacacct gatttaaata
tgactcgcca aaaagcagca 2820actcctgtca ccatcaatgt cacgattaat ggcaaatggc
aatctgctga taaaaatagt 2880gaagtgaaat atcaggtttc tggtgataac actgaactga
cgtttacgag ttactttggt 2940attccacaag aaatcaaact ctcgccactc ccttga
297685991PRTArtificialSynthetic polypeptide, HIV-1
TAT chondroitinase ABCI-N(delta)20 fusion polypeptide 85Gly Arg Lys
Lys Arg Arg Gln Arg Arg Arg Pro Pro Gln Cys Ala Gln 1 5
10 15 Asn Asn Pro Leu Ala Asp Phe Ser
Ser Asp Lys Asn Ser Ile Leu Thr 20 25
30 Leu Ser Asp Lys Arg Ser Ile Met Gly Asn Gln Ser Leu
Leu Trp Lys 35 40 45
Trp Lys Gly Gly Ser Ser Phe Thr Leu His Lys Lys Leu Ile Val Pro 50
55 60 Thr Asp Lys Glu
Ala Ser Lys Ala Trp Gly Arg Ser Ser Thr Pro Val 65 70
75 80 Phe Ser Phe Trp Leu Tyr Asn Glu Lys
Pro Ile Asp Gly Tyr Leu Thr 85 90
95 Ile Asp Phe Gly Glu Lys Leu Ile Ser Thr Ser Glu Ala Gln
Ala Gly 100 105 110
Phe Lys Val Lys Leu Asp Phe Thr Gly Trp Arg Thr Val Gly Val Ser
115 120 125 Leu Asn Asn Asp
Leu Glu Asn Arg Glu Met Thr Leu Asn Ala Thr Asn 130
135 140 Thr Ser Ser Asp Gly Thr Gln Asp
Ser Ile Gly Arg Ser Leu Gly Ala 145 150
155 160 Lys Val Asp Ser Ile Arg Phe Lys Ala Pro Ser Asn
Val Ser Gln Gly 165 170
175 Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser Val Asp Asp Ala Arg Tyr
180 185 190 Gln Trp Ser
Asp Tyr Gln Val Lys Thr Arg Leu Ser Glu Pro Glu Ile 195
200 205 Gln Phe His Asn Val Lys Pro Gln
Leu Pro Val Thr Pro Glu Asn Leu 210 215
220 Ala Ala Ile Asp Leu Ile Arg Gln Arg Leu Ile Asn Glu
Phe Val Gly 225 230 235
240 Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu Glu Asn Ile Ser Lys Leu
245 250 255 Lys Ser Asp Phe
Asp Ala Leu Asn Thr His Thr Leu Ala Asn Gly Gly 260
265 270 Thr Gln Gly Arg His Leu Ile Thr Asp
Lys Gln Ile Ile Ile Tyr Gln 275 280
285 Pro Glu Asn Leu Asn Ser Gln Asp Lys Gln Leu Phe Asp Asn
Tyr Val 290 295 300
Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe Asn Ile Ser Arg Ala Tyr 305
310 315 320 Val Leu Glu Lys Asp
Pro Thr Gln Lys Ala Gln Leu Lys Gln Met Tyr 325
330 335 Leu Leu Met Thr Lys His Leu Leu Asp Gln
Gly Phe Val Lys Gly Ser 340 345
350 Ala Leu Val Thr Thr His His Trp Gly Tyr Ser Ser Arg Trp Trp
Tyr 355 360 365 Ile
Ser Thr Leu Leu Met Ser Asp Ala Leu Lys Glu Ala Asn Leu Gln 370
375 380 Thr Gln Val Tyr Asp Ser
Leu Leu Trp Tyr Ser Arg Glu Phe Lys Ser 385 390
395 400 Ser Phe Asp Met Lys Val Ser Ala Asp Ser Ser
Asp Leu Asp Tyr Phe 405 410
415 Asn Thr Leu Ser Arg Gln His Leu Ala Leu Leu Leu Leu Glu Pro Asp
420 425 430 Asp Gln
Lys Arg Ile Asn Leu Val Asn Thr Phe Ser His Tyr Ile Thr 435
440 445 Gly Ala Leu Thr Gln Val Pro
Pro Gly Gly Lys Asp Gly Leu Arg Pro 450 455
460 Asp Gly Thr Ala Trp Arg His Glu Gly Asn Tyr Pro
Gly Tyr Ser Phe 465 470 475
480 Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile Tyr Leu Leu Arg Asp Thr
485 490 495 Pro Phe Ser
Val Gly Glu Ser Gly Trp Asn Ser Leu Lys Lys Ala Met 500
505 510 Val Ser Ala Trp Ile Tyr Ser Asn
Pro Glu Val Gly Leu Pro Leu Ala 515 520
525 Gly Arg His Pro Leu Asn Ser Pro Ser Leu Lys Ser Val
Ala Gln Gly 530 535 540
Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser Ser Pro Asp Lys Thr Leu 545
550 555 560 Ala Ser Ile Tyr
Leu Ala Ile Ser Asp Lys Thr Gln Asn Glu Ser Thr 565
570 575 Ala Ile Phe Gly Glu Thr Ile Thr Pro
Ala Ser Leu Pro Gln Gly Phe 580 585
590 Tyr Ala Phe Asn Gly Gly Ala Phe Gly Ile His Arg Trp Gln
Asp Lys 595 600 605
Met Val Thr Leu Lys Ala Tyr Asn Thr Asn Val Trp Ser Ser Glu Ile 610
615 620 Tyr Asn Lys Asp Asn
Arg Tyr Gly Arg Tyr Gln Ser His Gly Val Ala 625 630
635 640 Gln Ile Val Ser Asn Gly Ser Gln Leu Ser
Gln Gly Tyr Gln Gln Glu 645 650
655 Gly Trp Asp Trp Asn Arg Met Pro Gly Ala Thr Thr Ile His Leu
Pro 660 665 670 Leu
Lys Asp Leu Asp Ser Pro Lys Pro His Thr Leu Met Gln Arg Gly 675
680 685 Glu Arg Gly Phe Ser Gly
Thr Ser Ser Leu Glu Gly Gln Tyr Gly Met 690 695
700 Met Ala Phe Asp Leu Ile Tyr Pro Ala Asn Leu
Glu Arg Phe Asp Pro 705 710 715
720 Asn Phe Thr Ala Lys Lys Ser Val Leu Ala Ala Asp Asn His Leu Ile
725 730 735 Phe Ile
Gly Ser Asn Ile Asn Ser Ser Asp Lys Asn Lys Asn Val Glu 740
745 750 Thr Thr Leu Phe Gln His Ala
Ile Thr Pro Thr Leu Asn Thr Leu Trp 755 760
765 Ile Asn Gly Gln Lys Ile Glu Asn Met Pro Tyr Gln
Thr Thr Leu Gln 770 775 780
Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly Asn Gly Tyr Leu Ile Thr 785
790 795 800 Gln Ala Glu
Lys Val Asn Val Ser Arg Gln His Gln Val Ser Ala Glu 805
810 815 Asn Lys Asn Arg Gln Pro Thr Glu
Gly Asn Phe Ser Ser Ala Trp Ile 820 825
830 Asp His Ser Thr Arg Pro Lys Asp Ala Ser Tyr Glu Tyr
Met Val Phe 835 840 845
Leu Asp Ala Thr Pro Glu Lys Met Gly Glu Met Ala Gln Lys Phe Arg 850
855 860 Glu Asn Asn Gly
Leu Tyr Gln Val Leu Arg Lys Asp Lys Asp Val His 865 870
875 880 Ile Ile Leu Asp Lys Leu Ser Asn Val
Thr Gly Tyr Ala Phe Tyr Gln 885 890
895 Pro Ala Ser Ile Glu Asp Lys Trp Ile Lys Lys Val Asn Lys
Pro Ala 900 905 910
Ile Val Met Thr His Arg Gln Lys Asp Thr Leu Ile Val Ser Ala Val
915 920 925 Thr Pro Asp Leu
Asn Met Thr Arg Gln Lys Ala Ala Thr Pro Val Thr 930
935 940 Ile Asn Val Thr Ile Asn Gly Lys
Trp Gln Ser Ala Asp Lys Asn Ser 945 950
955 960 Glu Val Lys Tyr Gln Val Ser Gly Asp Asn Thr Glu
Leu Thr Phe Thr 965 970
975 Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys Leu Ser Pro Leu Pro
980 985 990
862856DNAArtificialSynthetic polynucleotide, HIV-1 TAT-
Chondroitinase ABC I N(delta)60 Nucleic Acid 86ggtcgtaaaa agcgtcgtca
acgtcgtcgt cctcctcaat gctttacttt acataaaaaa 60ctgattgtcc ccaccgataa
agaagcatct aaagcatggg gacgctcatc cacccccgtt 120ttctcatttt ggctttacaa
tgaaaaaccg attgatggtt atcttactat cgatttcgga 180gaaaaactca tttcaaccag
tgaggctcag gcaggcttta aagtaaaatt agatttcact 240ggctggcgta ctgtgggagt
ctctttaaat aacgatcttg aaaatcgaga gatgacctta 300aatgcaacca atacctcctc
tgatggtact caagacagca ttgggcgttc tttaggtgct 360aaagtcgata gtattcgttt
taaagcgcct tctaatgtga gtcagggtga aatctatatc 420gaccgtatta tgttttctgt
cgatgatgct cgctaccaat ggtctgatta tcaagtaaaa 480actcgcttat cagaacctga
aattcaattt cacaacgtaa agccacaact acctgtaaca 540cctgaaaatt tagcggccat
tgatcttatt cgccaacgtc taattaatga atttgtcgga 600ggtgaaaaag agacaaacct
cgcattagaa gagaatatca gcaaattaaa aagtgatttc 660gatgctctta atactcacac
tttagcaaat ggtggaacgc aaggcagaca tctgatcact 720gataaacaaa tcattattta
tcaaccagag aatcttaact ctcaagataa acaactattt 780gataattatg ttattttagg
taattacacg acattaatgt ttaatattag ccgtgcttat 840gtgctggaaa aagatcccac
acaaaaggcg caactaaagc agatgtactt attaatgaca 900aagcatttat tagatcaagg
ctttgttaaa gggagtgctt tagtgacaac ccatcactgg 960ggatacagtt ctcgttggtg
gtatatttcc acgttattaa tgtctgatgc actaaaagaa 1020gcgaacctac aaactcaagt
ttatgattca ttactgtggt attcacgtga gtttaaaagt 1080agttttgata tgaaagtaag
tgctgatagc tctgatctag attatttcaa taccttatct 1140cgccaacatt tagccttatt
actactagag cctgatgatc aaaagcgtat caacttagtt 1200aatactttca gccattatat
cactggcgca ttaacgcaag tgccaccggg tggtaaagat 1260ggtttacgcc ctgatggtac
agcatggcga catgaaggca actatccggg ctactctttc 1320ccagccttta aaaatgcctc
tcagcttatt tatttattac gcgatacacc attttcagtg 1380ggtgaaagtg gttggaatag
cctgaaaaaa gcgatggttt cagcgtggat ctacagtaat 1440ccagaagttg gattaccgct
tgcaggaaga caccctctta actcaccttc gttaaaatca 1500gtcgctcaag gctattactg
gcttgccatg tctgcaaaat catcgcctga taaaacactt 1560gcatctattt atcttgcgat
tagtgataaa acacaaaatg aatcaactgc tatttttgga 1620gaaactatta caccagcgtc
tttacctcaa ggtttctatg cctttaatgg cggtgctttt 1680ggtattcatc gttggcaaga
taaaatggtg acactgaaag cttataacac caatgtttgg 1740tcatctgaaa tttataacaa
agataaccgt tatggccgtt accaaagtca tggtgtcgct 1800caaatagtga gtaatggctc
gcagctttca cagggctatc agcaagaagg ttgggattgg 1860aatagaatgc caggggcaac
cactatccac cttcctctta aagacttaga cagtcctaaa 1920cctcatacct taatgcaacg
tggagagcgt ggatttagcg gaacatcatc ccttgaaggt 1980caatatggca tgatggcatt
cgatcttatt tatcccgcca atcttgagcg ttttgatcct 2040aatttcactg cgaaaaagag
tgtattagcc gctgataatc acttaatttt tattggtagc 2100aatataaata gtagtgataa
aaataaaaat gttgaaacga ccttattcca acatgccatt 2160actccaacat taaataccct
ttggattaat ggacaaaaga tagaaaacat gccttatcaa 2220acaacacttc aacaaggtga
ttggttaatt gatagcaatg gcaatggtta cttaattact 2280caagcagaaa aagtaaatgt
aagtcgccaa catcaggttt cagcggaaaa taaaaatcgc 2340caaccgacag aaggaaactt
tagctcggca tggatcgatc acagcactcg ccccaaagat 2400gccagttatg agtatatggt
ctttttagat gcgacacctg aaaaaatggg agagatggca 2460caaaaattcc gtgaaaataa
tgggttatat caggttcttc gtaaggataa agacgttcat 2520attattctcg ataaactcag
caatgtaacg ggatatgcct tttatcagcc agcatcaatt 2580gaagacaaat ggatcaaaaa
ggttaataaa cctgcaattg tgatgactca tcgacaaaaa 2640gacactctta ttgtcagtgc
agttacacct gatttaaata tgactcgcca aaaagcagca 2700actcctgtca ccatcaatgt
cacgattaat ggcaaatggc aatctgctga taaaaatagt 2760gaagtgaaat atcaggtttc
tggtgataac actgaactga cgtttacgag ttactttggt 2820attccacaag aaatcaaact
ctcgccactc ccttga
285687951PRTArtificialSynthetic polypeptide, HIV-1 TAT chondroitinase
ABCI-N(delta)60 fusion polypeptide 87Gly Arg Lys Lys Arg Arg Gln Arg Arg
Arg Pro Pro Gln Cys Phe Thr 1 5 10
15 Leu His Lys Lys Leu Ile Val Pro Thr Asp Lys Glu Ala Ser
Lys Ala 20 25 30
Trp Gly Arg Ser Ser Thr Pro Val Phe Ser Phe Trp Leu Tyr Asn Glu
35 40 45 Lys Pro Ile Asp
Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile 50
55 60 Ser Thr Ser Glu Ala Gln Ala Gly
Phe Lys Val Lys Leu Asp Phe Thr 65 70
75 80 Gly Trp Arg Thr Val Gly Val Ser Leu Asn Asn Asp
Leu Glu Asn Arg 85 90
95 Glu Met Thr Leu Asn Ala Thr Asn Thr Ser Ser Asp Gly Thr Gln Asp
100 105 110 Ser Ile Gly
Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg Phe Lys 115
120 125 Ala Pro Ser Asn Val Ser Gln Gly
Glu Ile Tyr Ile Asp Arg Ile Met 130 135
140 Phe Ser Val Asp Asp Ala Arg Tyr Gln Trp Ser Asp Tyr
Gln Val Lys 145 150 155
160 Thr Arg Leu Ser Glu Pro Glu Ile Gln Phe His Asn Val Lys Pro Gln
165 170 175 Leu Pro Val Thr
Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg Gln 180
185 190 Arg Leu Ile Asn Glu Phe Val Gly Gly
Glu Lys Glu Thr Asn Leu Ala 195 200
205 Leu Glu Glu Asn Ile Ser Lys Leu Lys Ser Asp Phe Asp Ala
Leu Asn 210 215 220
Thr His Thr Leu Ala Asn Gly Gly Thr Gln Gly Arg His Leu Ile Thr 225
230 235 240 Asp Lys Gln Ile Ile
Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp 245
250 255 Lys Gln Leu Phe Asp Asn Tyr Val Ile Leu
Gly Asn Tyr Thr Thr Leu 260 265
270 Met Phe Asn Ile Ser Arg Ala Tyr Val Leu Glu Lys Asp Pro Thr
Gln 275 280 285 Lys
Ala Gln Leu Lys Gln Met Tyr Leu Leu Met Thr Lys His Leu Leu 290
295 300 Asp Gln Gly Phe Val Lys
Gly Ser Ala Leu Val Thr Thr His His Trp 305 310
315 320 Gly Tyr Ser Ser Arg Trp Trp Tyr Ile Ser Thr
Leu Leu Met Ser Asp 325 330
335 Ala Leu Lys Glu Ala Asn Leu Gln Thr Gln Val Tyr Asp Ser Leu Leu
340 345 350 Trp Tyr
Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys Val Ser Ala 355
360 365 Asp Ser Ser Asp Leu Asp Tyr
Phe Asn Thr Leu Ser Arg Gln His Leu 370 375
380 Ala Leu Leu Leu Leu Glu Pro Asp Asp Gln Lys Arg
Ile Asn Leu Val 385 390 395
400 Asn Thr Phe Ser His Tyr Ile Thr Gly Ala Leu Thr Gln Val Pro Pro
405 410 415 Gly Gly Lys
Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg His Glu 420
425 430 Gly Asn Tyr Pro Gly Tyr Ser Phe
Pro Ala Phe Lys Asn Ala Ser Gln 435 440
445 Leu Ile Tyr Leu Leu Arg Asp Thr Pro Phe Ser Val Gly
Glu Ser Gly 450 455 460
Trp Asn Ser Leu Lys Lys Ala Met Val Ser Ala Trp Ile Tyr Ser Asn 465
470 475 480 Pro Glu Val Gly
Leu Pro Leu Ala Gly Arg His Pro Leu Asn Ser Pro 485
490 495 Ser Leu Lys Ser Val Ala Gln Gly Tyr
Tyr Trp Leu Ala Met Ser Ala 500 505
510 Lys Ser Ser Pro Asp Lys Thr Leu Ala Ser Ile Tyr Leu Ala
Ile Ser 515 520 525
Asp Lys Thr Gln Asn Glu Ser Thr Ala Ile Phe Gly Glu Thr Ile Thr 530
535 540 Pro Ala Ser Leu Pro
Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe 545 550
555 560 Gly Ile His Arg Trp Gln Asp Lys Met Val
Thr Leu Lys Ala Tyr Asn 565 570
575 Thr Asn Val Trp Ser Ser Glu Ile Tyr Asn Lys Asp Asn Arg Tyr
Gly 580 585 590 Arg
Tyr Gln Ser His Gly Val Ala Gln Ile Val Ser Asn Gly Ser Gln 595
600 605 Leu Ser Gln Gly Tyr Gln
Gln Glu Gly Trp Asp Trp Asn Arg Met Pro 610 615
620 Gly Ala Thr Thr Ile His Leu Pro Leu Lys Asp
Leu Asp Ser Pro Lys 625 630 635
640 Pro His Thr Leu Met Gln Arg Gly Glu Arg Gly Phe Ser Gly Thr Ser
645 650 655 Ser Leu
Glu Gly Gln Tyr Gly Met Met Ala Phe Asp Leu Ile Tyr Pro 660
665 670 Ala Asn Leu Glu Arg Phe Asp
Pro Asn Phe Thr Ala Lys Lys Ser Val 675 680
685 Leu Ala Ala Asp Asn His Leu Ile Phe Ile Gly Ser
Asn Ile Asn Ser 690 695 700
Ser Asp Lys Asn Lys Asn Val Glu Thr Thr Leu Phe Gln His Ala Ile 705
710 715 720 Thr Pro Thr
Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile Glu Asn 725
730 735 Met Pro Tyr Gln Thr Thr Leu Gln
Gln Gly Asp Trp Leu Ile Asp Ser 740 745
750 Asn Gly Asn Gly Tyr Leu Ile Thr Gln Ala Glu Lys Val
Asn Val Ser 755 760 765
Arg Gln His Gln Val Ser Ala Glu Asn Lys Asn Arg Gln Pro Thr Glu 770
775 780 Gly Asn Phe Ser
Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys Asp 785 790
795 800 Ala Ser Tyr Glu Tyr Met Val Phe Leu
Asp Ala Thr Pro Glu Lys Met 805 810
815 Gly Glu Met Ala Gln Lys Phe Arg Glu Asn Asn Gly Leu Tyr
Gln Val 820 825 830
Leu Arg Lys Asp Lys Asp Val His Ile Ile Leu Asp Lys Leu Ser Asn
835 840 845 Val Thr Gly Tyr
Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp 850
855 860 Ile Lys Lys Val Asn Lys Pro Ala
Ile Val Met Thr His Arg Gln Lys 865 870
875 880 Asp Thr Leu Ile Val Ser Ala Val Thr Pro Asp Leu
Asn Met Thr Arg 885 890
895 Gln Lys Ala Ala Thr Pro Val Thr Ile Asn Val Thr Ile Asn Gly Lys
900 905 910 Trp Gln Ser
Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val Ser Gly 915
920 925 Asp Asn Thr Glu Leu Thr Phe Thr
Ser Tyr Phe Gly Ile Pro Gln Glu 930 935
940 Ile Lys Leu Ser Pro Leu Pro 945 950
883036DNAArtificialSynthetic polynucleotide, C terminal HIV-1
TAT-Chondroitinase ABC I Nucleic acid 88gccaccagca atcctgcatt tgatcctaaa
aatctgatgc agtcagaaat ttaccatttt 60gcacaaaata acccattagc agacttctca
tcagataaaa actcaatact aacgttatct 120gataaacgta gcattatggg aaaccaatct
cttttatgga aatggaaagg tggtagtagc 180tttactttac ataaaaaact gattgtcccc
accgataaag aagcatctaa agcatgggga 240cgctcatcca cccccgtttt ctcattttgg
ctttacaatg aaaaaccgat tgatggttat 300cttactatcg atttcggaga aaaactcatt
tcaaccagtg aggctcaggc aggctttaaa 360gtaaaattag atttcactgg ctggcgtact
gtgggagtct ctttaaataa cgatcttgaa 420aatcgagaga tgaccttaaa tgcaaccaat
acctcctctg atggtactca agacagcatt 480gggcgttctt taggtgctaa agtcgatagt
attcgtttta aagcgccttc taatgtgagt 540cagggtgaaa tctatatcga ccgtattatg
ttttctgtcg atgatgctcg ctaccaatgg 600tctgattatc aagtaaaaac tcgcttatca
gaacctgaaa ttcaatttca caacgtaaag 660ccacaactac ctgtaacacc tgaaaattta
gcggccattg atcttattcg ccaacgtcta 720attaatgaat ttgtcggagg tgaaaaagag
acaaacctcg cattagaaga gaatatcagc 780aaattaaaaa gtgatttcga tgctcttaat
actcacactt tagcaaatgg tggaacgcaa 840ggcagacatc tgatcactga taaacaaatc
attatttatc aaccagagaa tcttaactct 900caagataaac aactatttga taattatgtt
attttaggta attacacgac attaatgttt 960aatattagcc gtgcttatgt gctggaaaaa
gatcccacac aaaaggcgca actaaagcag 1020atgtacttat taatgacaaa gcatttatta
gatcaaggct ttgttaaagg gagtgcttta 1080gtgacaaccc atcactgggg atacagttct
cgttggtggt atatttccac gttattaatg 1140tctgatgcac taaaagaagc gaacctacaa
actcaagttt atgattcatt actgtggtat 1200tcacgtgagt ttaaaagtag ttttgatatg
aaagtaagtg ctgatagctc tgatctagat 1260tatttcaata ccttatctcg ccaacattta
gccttattac tactagagcc tgatgatcaa 1320aagcgtatca acttagttaa tactttcagc
cattatatca ctggcgcatt aacgcaagtg 1380ccaccgggtg gtaaagatgg tttacgccct
gatggtacag catggcgaca tgaaggcaac 1440tatccgggct actctttccc agcctttaaa
aatgcctctc agcttattta tttattacgc 1500gatacaccat tttcagtggg tgaaagtggt
tggaatagcc tgaaaaaagc gatggtttca 1560gcgtggatct acagtaatcc agaagttgga
ttaccgcttg caggaagaca ccctcttaac 1620tcaccttcgt taaaatcagt cgctcaaggc
tattactggc ttgccatgtc tgcaaaatca 1680tcgcctgata aaacacttgc atctatttat
cttgcgatta gtgataaaac acaaaatgaa 1740tcaactgcta tttttggaga aactattaca
ccagcgtctt tacctcaagg tttctatgcc 1800tttaatggcg gtgcttttgg tattcatcgt
tggcaagata aaatggtgac actgaaagct 1860tataacacca atgtttggtc atctgaaatt
tataacaaag ataaccgtta tggccgttac 1920caaagtcatg gtgtcgctca aatagtgagt
aatggctcgc agctttcaca gggctatcag 1980caagaaggtt gggattggaa tagaatgcca
ggggcaacca ctatccacct tcctcttaaa 2040gacttagaca gtcctaaacc tcatacctta
atgcaacgtg gagagcgtgg atttagcgga 2100acatcatccc ttgaaggtca atatggcatg
atggcattcg atcttattta tcccgccaat 2160cttgagcgtt ttgatcctaa tttcactgcg
aaaaagagtg tattagccgc tgataatcac 2220ttaattttta ttggtagcaa tataaatagt
agtgataaaa ataaaaatgt tgaaacgacc 2280ttattccaac atgccattac tccaacatta
aatacccttt ggattaatgg acaaaagata 2340gaaaacatgc cttatcaaac aacacttcaa
caaggtgatt ggttaattga tagcaatggc 2400aatggttact taattactca agcagaaaaa
gtaaatgtaa gtcgccaaca tcaggtttca 2460gcggaaaata aaaatcgcca accgacagaa
ggaaacttta gctcggcatg gatcgatcac 2520agcactcgcc ccaaagatgc cagttatgag
tatatggtct ttttagatgc gacacctgaa 2580aaaatgggag agatggcaca aaaattccgt
gaaaataatg ggttatatca ggttcttcgt 2640aaggataaag acgttcatat tattctcgat
aaactcagca atgtaacggg atatgccttt 2700tatcagccag catcaattga agacaaatgg
atcaaaaagg ttaataaacc tgcaattgtg 2760atgactcatc gacaaaaaga cactcttatt
gtcagtgcag ttacacctga tttaaatatg 2820actcgccaaa aagcagcaac tcctgtcacc
atcaatgtca cgattaatgg caaatggcaa 2880tctgctgata aaaatagtga agtgaaatat
caggtttctg gtgataacac tgaactgacg 2940tttacgagtt actttggtat tccacaagaa
atcaaactct cgccactccc tggtcgtaaa 3000aagcgtcgtc aacgtcgtcg tcctcctcaa
tgctag 3036891011PRTArtificialSynthetic
polypeptide, C terminal HIV-1 TAT-Chondroitinase ABC I 89Ala Thr Ser
Asn Pro Ala Phe Asp Pro Lys Asn Leu Met Gln Ser Glu 1 5
10 15 Ile Tyr His Phe Ala Gln Asn Asn
Pro Leu Ala Asp Phe Ser Ser Asp 20 25
30 Lys Asn Ser Ile Leu Thr Leu Ser Asp Lys Arg Ser Ile
Met Gly Asn 35 40 45
Gln Ser Leu Leu Trp Lys Trp Lys Gly Gly Ser Ser Phe Thr Leu His 50
55 60 Lys Lys Leu Ile
Val Pro Thr Asp Lys Glu Ala Ser Lys Ala Trp Gly 65 70
75 80 Arg Ser Ser Thr Pro Val Phe Ser Phe
Trp Leu Tyr Asn Glu Lys Pro 85 90
95 Ile Asp Gly Tyr Leu Thr Ile Asp Phe Gly Glu Lys Leu Ile
Ser Thr 100 105 110
Ser Glu Ala Gln Ala Gly Phe Lys Val Lys Leu Asp Phe Thr Gly Trp
115 120 125 Arg Thr Val Gly
Val Ser Leu Asn Asn Asp Leu Glu Asn Arg Glu Met 130
135 140 Thr Leu Asn Ala Thr Asn Thr Ser
Ser Asp Gly Thr Gln Asp Ser Ile 145 150
155 160 Gly Arg Ser Leu Gly Ala Lys Val Asp Ser Ile Arg
Phe Lys Ala Pro 165 170
175 Ser Asn Val Ser Gln Gly Glu Ile Tyr Ile Asp Arg Ile Met Phe Ser
180 185 190 Val Asp Asp
Ala Arg Tyr Gln Trp Ser Asp Tyr Gln Val Lys Thr Arg 195
200 205 Leu Ser Glu Pro Glu Ile Gln Phe
His Asn Val Lys Pro Gln Leu Pro 210 215
220 Val Thr Pro Glu Asn Leu Ala Ala Ile Asp Leu Ile Arg
Gln Arg Leu 225 230 235
240 Ile Asn Glu Phe Val Gly Gly Glu Lys Glu Thr Asn Leu Ala Leu Glu
245 250 255 Glu Asn Ile Ser
Lys Leu Lys Ser Asp Phe Asp Ala Leu Asn Thr His 260
265 270 Thr Leu Ala Asn Gly Gly Thr Gln Gly
Arg His Leu Ile Thr Asp Lys 275 280
285 Gln Ile Ile Ile Tyr Gln Pro Glu Asn Leu Asn Ser Gln Asp
Lys Gln 290 295 300
Leu Phe Asp Asn Tyr Val Ile Leu Gly Asn Tyr Thr Thr Leu Met Phe 305
310 315 320 Asn Ile Ser Arg Ala
Tyr Val Leu Glu Lys Asp Pro Thr Gln Lys Ala 325
330 335 Gln Leu Lys Gln Met Tyr Leu Leu Met Thr
Lys His Leu Leu Asp Gln 340 345
350 Gly Phe Val Lys Gly Ser Ala Leu Val Thr Thr His His Trp Gly
Tyr 355 360 365 Ser
Ser Arg Trp Trp Tyr Ile Ser Thr Leu Leu Met Ser Asp Ala Leu 370
375 380 Lys Glu Ala Asn Leu Gln
Thr Gln Val Tyr Asp Ser Leu Leu Trp Tyr 385 390
395 400 Ser Arg Glu Phe Lys Ser Ser Phe Asp Met Lys
Val Ser Ala Asp Ser 405 410
415 Ser Asp Leu Asp Tyr Phe Asn Thr Leu Ser Arg Gln His Leu Ala Leu
420 425 430 Leu Leu
Leu Glu Pro Asp Asp Gln Lys Arg Ile Asn Leu Val Asn Thr 435
440 445 Phe Ser His Tyr Ile Thr Gly
Ala Leu Thr Gln Val Pro Pro Gly Gly 450 455
460 Lys Asp Gly Leu Arg Pro Asp Gly Thr Ala Trp Arg
His Glu Gly Asn 465 470 475
480 Tyr Pro Gly Tyr Ser Phe Pro Ala Phe Lys Asn Ala Ser Gln Leu Ile
485 490 495 Tyr Leu Leu
Arg Asp Thr Pro Phe Ser Val Gly Glu Ser Gly Trp Asn 500
505 510 Ser Leu Lys Lys Ala Met Val Ser
Ala Trp Ile Tyr Ser Asn Pro Glu 515 520
525 Val Gly Leu Pro Leu Ala Gly Arg His Pro Leu Asn Ser
Pro Ser Leu 530 535 540
Lys Ser Val Ala Gln Gly Tyr Tyr Trp Leu Ala Met Ser Ala Lys Ser 545
550 555 560 Ser Pro Asp Lys
Thr Leu Ala Ser Ile Tyr Leu Ala Ile Ser Asp Lys 565
570 575 Thr Gln Asn Glu Ser Thr Ala Ile Phe
Gly Glu Thr Ile Thr Pro Ala 580 585
590 Ser Leu Pro Gln Gly Phe Tyr Ala Phe Asn Gly Gly Ala Phe
Gly Ile 595 600 605
His Arg Trp Gln Asp Lys Met Val Thr Leu Lys Ala Tyr Asn Thr Asn 610
615 620 Val Trp Ser Ser Glu
Ile Tyr Asn Lys Asp Asn Arg Tyr Gly Arg Tyr 625 630
635 640 Gln Ser His Gly Val Ala Gln Ile Val Ser
Asn Gly Ser Gln Leu Ser 645 650
655 Gln Gly Tyr Gln Gln Glu Gly Trp Asp Trp Asn Arg Met Pro Gly
Ala 660 665 670 Thr
Thr Ile His Leu Pro Leu Lys Asp Leu Asp Ser Pro Lys Pro His 675
680 685 Thr Leu Met Gln Arg Gly
Glu Arg Gly Phe Ser Gly Thr Ser Ser Leu 690 695
700 Glu Gly Gln Tyr Gly Met Met Ala Phe Asp Leu
Ile Tyr Pro Ala Asn 705 710 715
720 Leu Glu Arg Phe Asp Pro Asn Phe Thr Ala Lys Lys Ser Val Leu Ala
725 730 735 Ala Asp
Asn His Leu Ile Phe Ile Gly Ser Asn Ile Asn Ser Ser Asp 740
745 750 Lys Asn Lys Asn Val Glu Thr
Thr Leu Phe Gln His Ala Ile Thr Pro 755 760
765 Thr Leu Asn Thr Leu Trp Ile Asn Gly Gln Lys Ile
Glu Asn Met Pro 770 775 780
Tyr Gln Thr Thr Leu Gln Gln Gly Asp Trp Leu Ile Asp Ser Asn Gly 785
790 795 800 Asn Gly Tyr
Leu Ile Thr Gln Ala Glu Lys Val Asn Val Ser Arg Gln 805
810 815 His Gln Val Ser Ala Glu Asn Lys
Asn Arg Gln Pro Thr Glu Gly Asn 820 825
830 Phe Ser Ser Ala Trp Ile Asp His Ser Thr Arg Pro Lys
Asp Ala Ser 835 840 845
Tyr Glu Tyr Met Val Phe Leu Asp Ala Thr Pro Glu Lys Met Gly Glu 850
855 860 Met Ala Gln Lys
Phe Arg Glu Asn Asn Gly Leu Tyr Gln Val Leu Arg 865 870
875 880 Lys Asp Lys Asp Val His Ile Ile Leu
Asp Lys Leu Ser Asn Val Thr 885 890
895 Gly Tyr Ala Phe Tyr Gln Pro Ala Ser Ile Glu Asp Lys Trp
Ile Lys 900 905 910
Lys Val Asn Lys Pro Ala Ile Val Met Thr His Arg Gln Lys Asp Thr
915 920 925 Leu Ile Val Ser
Ala Val Thr Pro Asp Leu Asn Met Thr Arg Gln Lys 930
935 940 Ala Ala Thr Pro Val Thr Ile Asn
Val Thr Ile Asn Gly Lys Trp Gln 945 950
955 960 Ser Ala Asp Lys Asn Ser Glu Val Lys Tyr Gln Val
Ser Gly Asp Asn 965 970
975 Thr Glu Leu Thr Phe Thr Ser Tyr Phe Gly Ile Pro Gln Glu Ile Lys
980 985 990 Leu Ser Pro
Leu Pro Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Pro 995
1000 1005 Pro Gln Cys 1010
902814DNAArtificialSynthetic polynucleotide, chondroitinase
ABCI-N(delta)60 90tttactttac ataaaaaact gattgtcccc accgataaag aagcatctaa
agcatgggga 60cgctcatcca cccccgtttt ctcattttgg ctttacaatg aaaaaccgat
tgatggttat 120cttactatcg atttcggaga aaaactcatt tcaaccagtg aggctcaggc
aggctttaaa 180gtaaaattag atttcactgg ctggcgtact gtgggagtct ctttaaataa
cgatcttgaa 240aatcgagaga tgaccttaaa tgcaaccaat acctcctctg atggtactca
agacagcatt 300gggcgttctt taggtgctaa agtcgatagt attcgtttta aagcgccttc
taatgtgagt 360cagggtgaaa tctatatcga ccgtattatg ttttctgtcg atgatgctcg
ctaccaatgg 420tctgattatc aagtaaaaac tcgcttatca gaacctgaaa ttcaatttca
caacgtaaag 480ccacaactac ctgtaacacc tgaaaattta gcggccattg atcttattcg
ccaacgtcta 540attaatgaat ttgtcggagg tgaaaaagag acaaacctcg cattagaaga
gaatatcagc 600aaattaaaaa gtgatttcga tgctcttaat actcacactt tagcaaatgg
tggaacgcaa 660ggcagacatc tgatcactga taaacaaatc attatttatc aaccagagaa
tcttaactct 720caagataaac aactatttga taattatgtt attttaggta attacacgac
attaatgttt 780aatattagcc gtgcttatgt gctggaaaaa gatcccacac aaaaggcgca
actaaagcag 840atgtacttat taatgacaaa gcatttatta gatcaaggct ttgttaaagg
gagtgcttta 900gtgacaaccc atcactgggg atacagttct cgttggtggt atatttccac
gttattaatg 960tctgatgcac taaaagaagc gaacctacaa actcaagttt atgattcatt
actgtggtat 1020tcacgtgagt ttaaaagtag ttttgatatg aaagtaagtg ctgatagctc
tgatctagat 1080tatttcaata ccttatctcg ccaacattta gccttattac tactagagcc
tgatgatcaa 1140aagcgtatca acttagttaa tactttcagc cattatatca ctggcgcatt
aacgcaagtg 1200ccaccgggtg gtaaagatgg tttacgccct gatggtacag catggcgaca
tgaaggcaac 1260tatccgggct actctttccc agcctttaaa aatgcctctc agcttattta
tttattacgc 1320gatacaccat tttcagtggg tgaaagtggt tggaatagcc tgaaaaaagc
gatggtttca 1380gcgtggatct acagtaatcc agaagttgga ttaccgcttg caggaagaca
ccctcttaac 1440tcaccttcgt taaaatcagt cgctcaaggc tattactggc ttgccatgtc
tgcaaaatca 1500tcgcctgata aaacacttgc atctatttat cttgcgatta gtgataaaac
acaaaatgaa 1560tcaactgcta tttttggaga aactattaca ccagcgtctt tacctcaagg
tttctatgcc 1620tttaatggcg gtgcttttgg tattcatcgt tggcaagata aaatggtgac
actgaaagct 1680tataacacca atgtttggtc atctgaaatt tataacaaag ataaccgtta
tggccgttac 1740caaagtcatg gtgtcgctca aatagtgagt aatggctcgc agctttcaca
gggctatcag 1800caagaaggtt gggattggaa tagaatgcca ggggcaacca ctatccacct
tcctcttaaa 1860gacttagaca gtcctaaacc tcatacctta atgcaacgtg gagagcgtgg
atttagcgga 1920acatcatccc ttgaaggtca atatggcatg atggcattcg atcttattta
tcccgccaat 1980cttgagcgtt ttgatcctaa tttcactgcg aaaaagagtg tattagccgc
tgataatcac 2040ttaattttta ttggtagcaa tataaatagt agtgataaaa ataaaaatgt
tgaaacgacc 2100ttattccaac atgccattac tccaacatta aatacccttt ggattaatgg
acaaaagata 2160gaaaacatgc cttatcaaac aacacttcaa caaggtgatt ggttaattga
tagcaatggc 2220aatggttact taattactca agcagaaaaa gtaaatgtaa gtcgccaaca
tcaggtttca 2280gcggaaaata aaaatcgcca accgacagaa ggaaacttta gctcggcatg
gatcgatcac 2340agcactcgcc ccaaagatgc cagttatgag tatatggtct ttttagatgc
gacacctgaa 2400aaaatgggag agatggcaca aaaattccgt gaaaataatg ggttatatca
ggttcttcgt 2460aaggataaag acgttcatat tattctcgat aaactcagca atgtaacggg
atatgccttt 2520tatcagccag catcaattga agacaaatgg atcaaaaagg ttaataaacc
tgcaattgtg 2580atgactcatc gacaaaaaga cactcttatt gtcagtgcag ttacacctga
tttaaatatg 2640actcgccaaa aagcagcaac tcctgtcacc atcaatgtca cgattaatgg
caaatggcaa 2700tctgctgata aaaatagtga agtgaaatat caggtttctg gtgataacac
tgaactgacg 2760tttacgagtt actttggtat tccacaagaa atcaaactct cgccactccc
ttga 2814912934DNAArtificialSynthetic polynucleotide,
chondroitinase ABCI-N(delta)20 91gcacaaaata acccattagc agacttctca
tcagataaaa actcaatact aacgttatct 60gataaacgta gcattatggg aaaccaatct
cttttatgga aatggaaagg tggtagtagc 120tttactttac ataaaaaact gattgtcccc
accgataaag aagcatctaa agcatgggga 180cgctcatcca cccccgtttt ctcattttgg
ctttacaatg aaaaaccgat tgatggttat 240cttactatcg atttcggaga aaaactcatt
tcaaccagtg aggctcaggc aggctttaaa 300gtaaaattag atttcactgg ctggcgtact
gtgggagtct ctttaaataa cgatcttgaa 360aatcgagaga tgaccttaaa tgcaaccaat
acctcctctg atggtactca agacagcatt 420gggcgttctt taggtgctaa agtcgatagt
attcgtttta aagcgccttc taatgtgagt 480cagggtgaaa tctatatcga ccgtattatg
ttttctgtcg atgatgctcg ctaccaatgg 540tctgattatc aagtaaaaac tcgcttatca
gaacctgaaa ttcaatttca caacgtaaag 600ccacaactac ctgtaacacc tgaaaattta
gcggccattg atcttattcg ccaacgtcta 660attaatgaat ttgtcggagg tgaaaaagag
acaaacctcg cattagaaga gaatatcagc 720aaattaaaaa gtgatttcga tgctcttaat
actcacactt tagcaaatgg tggaacgcaa 780ggcagacatc tgatcactga taaacaaatc
attatttatc aaccagagaa tcttaactct 840caagataaac aactatttga taattatgtt
attttaggta attacacgac attaatgttt 900aatattagcc gtgcttatgt gctggaaaaa
gatcccacac aaaaggcgca actaaagcag 960atgtacttat taatgacaaa gcatttatta
gatcaaggct ttgttaaagg gagtgcttta 1020gtgacaaccc atcactgggg atacagttct
cgttggtggt atatttccac gttattaatg 1080tctgatgcac taaaagaagc gaacctacaa
actcaagttt atgattcatt actgtggtat 1140tcacgtgagt ttaaaagtag ttttgatatg
aaagtaagtg ctgatagctc tgatctagat 1200tatttcaata ccttatctcg ccaacattta
gccttattac tactagagcc tgatgatcaa 1260aagcgtatca acttagttaa tactttcagc
cattatatca ctggcgcatt aacgcaagtg 1320ccaccgggtg gtaaagatgg tttacgccct
gatggtacag catggcgaca tgaaggcaac 1380tatccgggct actctttccc agcctttaaa
aatgcctctc agcttattta tttattacgc 1440gatacaccat tttcagtggg tgaaagtggt
tggaatagcc tgaaaaaagc gatggtttca 1500gcgtggatct acagtaatcc agaagttgga
ttaccgcttg caggaagaca ccctcttaac 1560tcaccttcgt taaaatcagt cgctcaaggc
tattactggc ttgccatgtc tgcaaaatca 1620tcgcctgata aaacacttgc atctatttat
cttgcgatta gtgataaaac acaaaatgaa 1680tcaactgcta tttttggaga aactattaca
ccagcgtctt tacctcaagg tttctatgcc 1740tttaatggcg gtgcttttgg tattcatcgt
tggcaagata aaatggtgac actgaaagct 1800tataacacca atgtttggtc atctgaaatt
tataacaaag ataaccgtta tggccgttac 1860caaagtcatg gtgtcgctca aatagtgagt
aatggctcgc agctttcaca gggctatcag 1920caagaaggtt gggattggaa tagaatgcca
ggggcaacca ctatccacct tcctcttaaa 1980gacttagaca gtcctaaacc tcatacctta
atgcaacgtg gagagcgtgg atttagcgga 2040acatcatccc ttgaaggtca atatggcatg
atggcattcg atcttattta tcccgccaat 2100cttgagcgtt ttgatcctaa tttcactgcg
aaaaagagtg tattagccgc tgataatcac 2160ttaattttta ttggtagcaa tataaatagt
agtgataaaa ataaaaatgt tgaaacgacc 2220ttattccaac atgccattac tccaacatta
aatacccttt ggattaatgg acaaaagata 2280gaaaacatgc cttatcaaac aacacttcaa
caaggtgatt ggttaattga tagcaatggc 2340aatggttact taattactca agcagaaaaa
gtaaatgtaa gtcgccaaca tcaggtttca 2400gcggaaaata aaaatcgcca accgacagaa
ggaaacttta gctcggcatg gatcgatcac 2460agcactcgcc ccaaagatgc cagttatgag
tatatggtct ttttagatgc gacacctgaa 2520aaaatgggag agatggcaca aaaattccgt
gaaaataatg ggttatatca ggttcttcgt 2580aaggataaag acgttcatat tattctcgat
aaactcagca atgtaacggg atatgccttt 2640tatcagccag catcaattga agacaaatgg
atcaaaaagg ttaataaacc tgcaattgtg 2700atgactcatc gacaaaaaga cactcttatt
gtcagtgcag ttacacctga tttaaatatg 2760actcgccaaa aagcagcaac tcctgtcacc
atcaatgtca cgattaatgg caaatggcaa 2820tctgctgata aaaatagtga agtgaaatat
caggtttctg gtgataacac tgaactgacg 2880tttacgagtt actttggtat tccacaagaa
atcaaactct cgccactccc ttga 29349251DNAArtificialOligonucleotide
used for cloning TAT-ABC I 92tatgtatggt cgtaaaaagc gtcgtcaacg tcgtcgtggt
ggtggtggtc a 519354DNAArtificialOTHER INFORMATION;
Oligonucleotide used for cloning TAT-ABC I 93tatgaccacc accaccacca
cgacgacgtt gacgacgctt tttacgacca taca
54947PRTArtificialArtificial Sequence, Gly-Ser linker 94Gly Gly Gly Gly
Ser Ser Ser 1 5 9527DNAArtificialPCR primer
95catatggcca ccagcmtcct gcatttg
279624DNAArtificialPCR primer 96ggatcctcaa gggagtggcg agag
24
User Contributions:
Comment about this patent or add new information about this topic: