Modulating robo: ligand interactions

Abstract

compositions for identifying agents which modulate the interaction of Robo and a Robo ligand and for modulating the interaction of Robo and a Robo ligand. The methods for identifying Robo:ligand modulators find particular application in commercial drug screens. These methods generally comprise (1) combining a Robo polypeptide, a Slit polypeptide and a candidate agent under conditions whereby, but for the presence of the agent, the Robo and Slit polypeptides engage in a first interaction, and (2) determining a second interaction of the Robo and Slit polypeptides in the presence of the agent, wherein a difference between the first and second interactions indicates that the aget modulates the interaction of the Robo and Slit polypeptides. The subject methods of modulating the interaction of Robo and a Robo ligand involve combining a Robo polypeptide, a Slit polypeptide and a modulator under conditions whereby, but for the presence of the modulator, the Robo and Slit polypeptides engage in a first interaction, whereby the Robo and Slit polypeptides engage in a second interaction different from the first interaction. In a particular embodiment, the modulator is dominant negative form of the Robo or Slit polypeptide.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001]This application is a continuation of U.S. application Ser. No. 11/022,546 filed Dec. 23, 2004, which is a division of U.S. application Ser. No. 10/289,776 filed Nov. 6, 2002, now U.S. Pat. No. 6,861,228, which is a continuation of U.S. application Ser. No. 09/922,600 filed Aug. 3, 2001, now abandoned, which is a continuation of U.S. application Ser. No. 09/540,245 filed Mar. 31, 2000, now U.S. Pat. No. 6,270,984, which is a division of U.S. application Ser. No. 09/191,647 filed Nov. 13, 1998, now U.S. Pat. No. 6,046,015 which claims the benefit of U.S. Provisional Application No. 60/081,057 filed Apr. 7, 1998 and U.S. Provisional Application No. 60/065,544 filed Nov. 14, 1997, all of which are incorporated herein by reference.

[0002]The research carried out in the subject application was supported in part by NIH grant NS18366. The government may have rights in any patent issuing on this application.

INTRODUCTION

[0003]1. Field of the Invention

[0004]The field of this invention is methods for modulating nerve cell function.

[0005]2. Background

[0006]In the developing CNS, most growth cones confront the midline at one or multiple times during their journey and make the decision of whether to cross or not to cross. This decision is not a static one but rather changes according to the growth cone's history. For example, in the Drosophila ventral nerve cord, about 10% of the interneurons project their axons only on their own side, in some cases extending near the midline without crossing it. The other 90% of the interneurons first project their axons across the midline and then turn to project longitudinally on the other side, often extending near the midline. These growth cones, having crossed the midline once, never cross it again, in spite of their close proximity to the midline and the many commissural axons crossing it. This decision to cross or not to cross is not unique to Drosophila but is common to a variety of midline structures in all bilaterally symmetric nervous systems.

[0007]What midline signals and growth cone receptors control whether growth cones do or do not cross the midline? After crossing once, what mechanism prevents these growth cones from crossing again? A related issue concerns the nature of the midline as an intermediate target. If so many growth cones find the midline such an attractive structure, why do they cross over it rather than linger? Why do they leave the midline?

[0008]One approach to find the genes encoding the components of such a system is to screen for mutations in which either too many or too few axons cross the midline. Such a large-scale mutant screen was previously conducted in Drosophila, and led to the identification of two key genes: commissureless (comm) and roundabout (robo) (Seeger et al., 1993; reviewed by Tear et al., 1993). In comm mutant embryos, commissural growth cones initially orient toward the midline but then fail to cross it and instead recoil and extend on their own side. robo mutant embryos, on the other hand, display the opposite phenotype in that too many axons cross the midline; many growth cones that normally extend only on their own side instead now project across the midline and axons that normally cross the midline only once instead appear to cross and recross multiple times (Seeger et al, 1993; present disclosure). Double mutants of comm and robo display a robo-like phenotype.

[0009]How do comm and robo function to control midline crossing? Neither the initial paper on these genes (Seeger et al., 1993) nor the cloning of comm (Tear et al., 1996) resolved this question. comm encodes a novel surface protein expressed on midline cells. In fact, the comm paper (Tear et al., 1996) ended with the hope that future work would ". . . help shed some light on the enigmatic function of Comm."

[0010]U.S. Ser. No. 08/971,172 (Robo, A Novel Family of Polypeptides and Nucleic Acids, by inventors: Corey S. Goodman, Thomas Kidd, Kevin J. Mitchell and Guy Tear) discloses the cloning and characterization of robo in various species including Drosophila; Robo polypeptides and polypeptide-encoding nucleic acids are also disclosed and their genbank accession numbers referenced in Kidd et al. (1998) Cell 92, 205-215. robo encodes a new class of guidance receptor with 5 immunoglobulin (Ig) domains, 3 fibronectin type III domains, a transmembrane domain, and a long cytoplasmic domain. Robo defines a new subfamily of Ig superfamily proteins that is highly conserved from fruit flies to mammals. The Robo ectodomains, and in particular the first two Ig domains, are highly conserved from fruit fly to human, while the cytoplasmic domains are more divergent. Nevertheless, the cytoplasmic domains contain three highly conserved short proline-rich motifs which may represent binding sites for SH3 or other binding domains in linker or signaling molecules.

[0011]For those axons that never cross the midline, Robo is expressed on their growth cones from the outset; for the majority of axons that do cross the midline, Robo is expressed at high levels on their growth cones only after they cross the midline. Transgenic rescue experiments in Drosophila reveal that Robo can function in a cell autonomous fashion, consistent with it functioning as a receptor. Thus, in Drosophila, Robo appears to function as the gatekeeper controlling midline crossing; growth cones expressing high levels of Robo are prevented from crossing the midline. Robo proteins in mammals function in a similar manner in controlling axon guidance.

[0012]U.S. Ser. No. 60/065,54 (Methods for Modulating Nerve Cell Function, by inventors: Corey S. Goodman, Thomas Kidd, Guy Tear, Claire Russell and Kevin Mitchell) discloses ectopic and overexpression studies revealing that Comm down-regulates Robo expression, demonstrating that Comm functions to suppress the Robo-mediated midline repulsion. These results show that the levels of Comm at the midline and Robo on growth cones are tightly intertwined and dynamically regulated to assure that only certain growth cones cross the midline, that those growth cones that cross do not linger at the midline, and that once they cross they never do so again.

Relevant Literature

Seeger, M., Tear, G., Ferres-Marco, D. and Goodman C. S. (1993) Neuron 10, 409 -426; Tear G., et al. (1996) Neuron 16, 501 -514; Rothberg et al. (1990) Genes Dev 4, 2169-2187; Kidd et al. (1998) Cell 92, 205-215.

SUMMARY OF THE INVENTION

[0013]The invention provides methods and compositions relating to vertebrate Slit1 and Slit2, collectively vertebrate Slit) polypeptides, related nucleic acids, polypeptide domains thereof having vertebrate Slit-specific structure and activity, and modulators of vertebrate Slit function. Vertebrate Slit polypeptides can regulate cell, especially nerve cell, function and morphology. The polypeptides may be produced recombinantly from transformed host cells from the subject vertebrate Slit polypeptide encoding nucleic acids or purified from mammalian cells. The invention provides isolated vertebrate Slit hybridization probes and primers capable of specifically hybridizing with natural vertebrate Slit genes, vertebrate Slit-specific binding agents such as specific antibodies, and methods of making and using the subject compositions in diagnosis (e.g. genetic hybridization screens for vertebrate Slit transcripts), therapy (e.g. to modulate nerve cell growth) and in the biopharmaceutical industry (e.g. as immunogens, reagents for isolating vertebrate Slit genes and polypeptides, reagents for screening chemical libraries for lead pharmacological agents, etc.).

[0014]The invention also provides methods and compositions for identifying agents which modulate the interaction of Robo and a Robo ligand and for modulating the interaction of Robo and a Robo ligand. The methods for identifying Robo:ligand modulators find particular application in commercial drug screens. These methods generally comprise (1) combining a Robo polypeptide, a Slit polypeptide and a candidate agent under conditions whereby, but for the presence of the agent, the Robo and Slit polypeptides engage in a first interaction, and (2) determining a second interaction of the Robo and Slit polypeptides in the presence of the agent, wherein a difference between the first and second interactions indicates that the aget modulates the interaction of the Robo and Slit polypeptides. The subject methods of modulating the interaction of Robo and a Robo ligand involve combining a Robo polypeptide, a Slit polypeptide and a modulator under conditions whereby, but for the presence of the modulator, the Robo and Slit polypeptides engage in a first interaction, whereby the Robo and Slit polypeptides engage in a second interaction different from the first interaction. In a particular embodiment, the modulator is dominant negative form of the Robo or Slit polypeptide.

DETAILED DESCRIPTION OF THE INVENTION

[0015]The subject methods include screens for agents which modulate Robo:ligand interactions and methods for modulating Robo:ligand interactions. Robo activation is found to regulate a wide variety of cell functions, including cell-cell interactions, cell mobility, morphology, etc. Slit polypeptides are disclosed as specific activators and inactivators of Robo polypeptides. Accordingly, the invention provides methods for modulating targeted cell function comprising the step of modulating Robo activation by contacting the cell with a modulator of a Robo:Slit interaction.

[0016]The targeted Robo polypeptide is generally naturally expressed on the targeted cells. The nucleotide sequences of exemplary natural cDNAs encoding drosophila 1, drosophila 2, C. elegans, human 1, human 2 and mouse 1 Robo polypeptides and their translates are described in Kidd et al. (1998) Cell 92, 205-215 and U.S. Ser. No. 08/971,172. The targeted Robo polypeptides comprise at least a functional Robo domain, which domain has Robo-specific amino acid sequence and binding specificity or function. Preferred Robo domains comprise at least 8, preferably at least 16, more preferably at least 32, most preferably at least 64 consecutive residues of a natural full length Robo. In a particular embodiment, the domains comprise one or more structural/functional Robo immunoglobulin, fibronectin or cytoplasmic motif domains described herein. The subject domains provide Robo-specific antigens and/or immunogens, especially when coupled to carrier proteins. For example, peptides corresponding to Robo- and human Robo-specific domains are covalently coupled to keyhole limpet antigen (KLH) and the conjugate is emulsified in Freunds complete adjuvant. Laboratory rabbits are immunized according to conventional protocol and bled. The presence of Robo-specific antibodies is assayed by solid phase immunosorbant assays using immobilized Robo polypeptides. Generic Robo-specific peptides are readily apparent as conserved regions in aligned Robo polypeptide sequences. In addition, species-specific antigenic and/or immunogenic peptides are readily apparent as diverged extracellular or cytosolic regions in alignments Human Robo-specific antibodies are characterized as uncross-reactive with non-human Robo polypeptides.

[0017]The subject domains provide Robo domain specific activity or function, such as Robo-specific cell, especially neuron modulating or modulating inhibitory activity, Robo-ligand-binding or binding inhibitory activity. Robo-specific activity or function may be determined by convenient in vitro, cell-based, or in vivo assays: e.g. in vitro binding assays, cell culture assays, in animals (e.g. gene therapy, transgenics, etc.), etc. The binding target may be a natural intracellular binding target, a Robo regulating protein or other regulator that directly modulates Robo activity or its localization; or non-natural binding target such as a specific immune protein such as an antibody, or a Robo specific agent such as those identified in screening assays such as described below. Robo-binding specificity may be assayed by binding equilibrium constants (usually at least about 10⁷ M^-1, preferably at least about 10⁸ M^-1, more preferably at least about 10⁹ M^-1), by the ability of the subject polypeptide to function as negative mutants in Robo-expressing cells, to elicit Robo specific antibody in a heterologous host (e.g a rodent or rabbit), etc.

[0018]Similarly, the Slit polypeptide is conveniently selected from Slit polypeptides which specifically activate or inhibit the activation of the Robo polypeptide. Exemplary suitable Slit polypeptides (a) comprises a vertebrate Slit sequence disclosed herein, especially human Slit-1 (SEQ ID NO:02), or a deletion mutant thereof which specifically modulates Robo expression or a sequence about 60-70%, preferably about 70-80%, more preferably about 80-90%, more preferably about 90-95%, most preferably about 95-99% similar to a vertebrate Slit sequence disclosed herein as determined by Best Fit analysis using default settings and is other than a natural drosophila Slit sequence, preferably other than a natural invertebrate Slit sequence, and/or (b) is encoded by a nucleic acid comprising a natural Slit encoding sequence (such as a natural human Slit-1 encoding sequence, SEQ ID NO:01) or a fragment thereof at least 36, preferably at least 72, more preferably at least 144, most preferably at least 288 nucleotides in length which specifically hybridizes thereto. Suitable deletion mutants are readily screened in Robo binding or activation assays as described herein. Preferred Slit domains/deletion mutants/fragments comprise at least 8, preferably at least 16, more preferably at least 32, most preferably at least 64 consecutive residues of a disclosed vertebrate Slit sequences and provide a Slit specific activity, such as Slit-specific antigenicity and/or immunogenicity, especially when coupled to carrier proteins as described above for Robo above. Suitable natural Slit encoding sequence fragments are of length sufficient to encode such Slit domains. In a particular embodiment, the Slit fragments comprise species specific fragments; such fragments are readily discerned from alignments of the disclosed sequences, see, e.g. shown as unboxed sequences in Tables 1 and 2. (See Appendix A) Exemplary such human Slit-1 immunogenic and/or antigenic peptides are shown in Table 3.

TABLE-US-00001 TABLE 3 Immunogenic human Slit-1 polypeptides eliciting Slit-1 specific rabbit polyclonal antibody: Slit polypeptide-KLH conjugates immunized per protocol described above. Slit Polypeptide Immunogenicity SEQ ID NO: 02, res. 1-10 +++ SEQ ID NO: 02, res. 29-41 +++ SEQ ID NO: 02, res. 75-87 +++ SEQ ID NO: 02, res. 92-109 +++ SEQ ID NO: 02, res. 132-141 +++ SEQ ID NO: 02, res. 192-205 +++ SEQ ID NO: 02, res. 258-269 +++ SEQ ID NO: 02, res. 295-311 +++ SEQ ID NO: 02, res. 316-330 +++ SEQ ID NO: 02, res. 373-382 +++ SEQ ID NO: 02, res. 403-422 +++ SEQ ID NO: 02, res. 474-485 +++ SEQ ID NO: 02, res. 561-576 +++ SEQ ID NO: 02, res. 683-697 +++ SEQ ID NO: 02, res. 768-777 +++ SEQ ID NO: 02, res. 798-813 +++ SEQ ID NO: 02, res. 882-894 +++ SEQ ID NO: 02, res. 934-946 +++ SEQ ID NO: 02, res. 1054-1067 +++ SEQ ID NO: 02, res. 1181-1192 +++ SEQ ID NO: 02, res. 1273-1299 +++ SEQ ID NO: 02, res. 1383-1397 +++ SEQ ID NO: 02, res. 1468-1477 +++ SEQ ID NO: 02, res. 1508-1517 +++

[0019]The subject domains provide Slit domain specific activity or function, such as Slit-specific cell, especially neuron modulating or modulating inhibitory activity, Slit-ligand-binding or binding inhibitory activity. Slit-specific activity or function may be determined by convenient in vitro, cell-based, or in vivo assays: e.g. in vitro binding assays, cell culture assays, in animals (e.g. gene therapy, transgenics, etc.), etc. The binding target may be a natural intracellular binding target, a Slit regulating protein or other regulator that directly modulates Slit activity or its localization; or non-natural binding target such as a specific immune protein such as an antibody, or a Slit specific agent such as those identified in screening assays such as described below. Slit-binding specificity may be assayed by binding equilibrium constants (usually at least about 10⁷ M^-1, preferably at least about 10⁸ M^-1, more preferably at least about 10⁹ M^-1), by the ability of the subject polypeptide to function as negative mutants in Slit-expressing cells, to elicit Slit specific antibody in a heterologous host (e.g a rodent or rabbit), etc.

[0020]In one embodiment, the Slit polypeptides are encoded by a nucleic acid comprising SEQ ID NO:01 or a fragment thereof which hybridizes with a full-length strand thereof, preferably under stringent conditions. Such nucleic acids comprise at least 36, preferably at least 72, more preferably at least 144 and most preferably at least 288 nucleotides of SEQ ID NO:01. Demonstrating specific hybridization generally requires stringent conditions, for example, hybridizing in a buffer comprising 30% formamide in 5×SSPE (0.18 M NaCl, 0.01 M NaPO₄, pH7.7, 0.001 M EDTA) buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE (Conditions 1); preferably hybridizing in a buffer comprising 50% formamide in 5×SSPE buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE buffer at 42° C. (Conditions II). Exemplary nucleic acids which hybridize with a strand of SEQ ID NO:01 are shown in Table 4.

TABLE-US-00002 TABLE 4 Exemplary nucleic acids which hybridize with a strand of SEQ ID NO: 01 under Conditions I and/or II. Slit Nucleic Acid Hybridization SEQ ID NO: 01, nucl. 1-47 + SEQ ID NO: 01, nucl. 58-99 + SEQ ID NO: 01, nucl. 95-138 + SEQ ID NO: 01, nucl. 181-220 + SEQ ID NO: 01, nucl. 261-299 + SEQ ID NO: 01, nucl. 274-315 + SEQ ID NO: 01, nucl. 351-389 + SEQ ID NO: 01, nucl. 450-593 + SEQ ID NO: 01, nucl. 524-546 + SEQ ID NO: 01, nucl. 561-608 + SEQ ID NO: 01, nucl. 689-727 + SEQ ID NO: 01, nucl. 708-737 + SEQ ID NO: 01, nucl. 738-801 + SEQ ID NO: 01, nucl. 805-854 + SEQ ID NO: 01, nucl. 855-907 + SEQ ID NO: 01, nucl. 910-953 + SEQ ID NO: 01, nucl. 1007-1059 + SEQ ID NO: 01, nucl. 1147-1163 + SEQ ID NO: 01, nucl. 1258-1279 + SEQ ID NO: 01, nucl. 1375-1389 + SEQ ID NO: 01, nucl. 1581-1595 + SEQ ID NO: 01, nucl. 1621-1639 + SEQ ID NO: 01, nucl. 1744-1755 + SEQ ID NO: 01, nucl. 1951-1969 + SEQ ID NO: 01, nucl. 2150-2163 + SEQ ID NO: 01, nucl. 2524-2546 + SEQ ID NO: 01, nucl. 2761-2780 + SEQ ID NO: 01, nucl. 2989-2999 + SEQ ID NO: 01, nucl. 3108-3117 + SEQ ID NO: 01, nucl. 3338-3351 + SEQ ID NO: 01, nucl. 3505-3514 + SEQ ID NO: 01, nucl. 3855-3867 + SEQ ID NO: 01, nucl. 4010-4025 + SEQ ID NO: 01, nucl. 4207-4219 + SEQ ID NO: 01, nucl. 4333-4345 + SEQ ID NO: 01, nucl. 4521-4529 +

[0021]A wide variety of cell types express Robo polypeptides subject to regulation by the disclosed methods, including many neuronal cells, transformed cells, infected (e.g. virus) cells, etc. Ascertaining Robo binding or activation is readily effected by binding assays or cells function assays as disclosed herein or in the cited copending applications. Accordingly, indications for the subject methods encompass a wide variety of cell types and function, including axon outgrowth, tumor cell invasion or migration, etc. The target cell may reside in culture or in situ, i.e. within the natural host. For in situ applications, the compositions are added to a retained physiological fluid such as blood or synovial fluid. For CNS administration, a variety of techniques are available for promoting transfer of the therapeutic across the blood brain barrier including disruption by surgery or injection, drugs which transiently open adhesion contact between CNS vasculature endothelial cells, and compounds which facilitate translocation through such cells. Slit polypeptides may also be amenable to direct injection or infusion, topical, intratracheal/nasal administration e.g. through aerosol, intraocularly, or within/on implants e.g. fibers e.g. collagen, osmotic pumps, grafts comprising appropriately transformed cells, etc. A particular method of administration involves coating, embedding or derivatizing fibers, such as collagen fibers, protein polymers, etc. with therapeutic polypeptides. Other useful approaches are described in Otto et al. (1989) J Neuroscience Research 22, 83-91 and Otto and Unsicker (1990) J Neuroscience 10, 1912-1921. Generally, the amount administered will be empirically determined, typically in the range of about 10 to 1000 μg/kg of the recipient and the concentration will generally be in the range of about 50 to 500 μg/ml in the dose administered. Other additives may be included, such as stabilizers, bactericides, etc. will be present in conventional amounts.

[0022]In one embodiment, the invention provides administering the subject Slit polypeptides in combination with a pharmaceutically acceptable excipient such as sterile saline or other medium, gelatin, an oil, etc. to form pharmaceutically acceptable compositions. The compositions and/or compounds may be administered alone or in combination with any convenient carrier, diluent, etc. and such administration may be provided in single or multiple dosages. Useful carriers include solid, semi-solid or liquid media including water and non-toxic organic solvents. In another embodiment, the invention provides the subject compounds in the form of a pro-drug, which can be metabolically converted to the subject compound by the recipient host. A wide variety of pro-drug formulations for polypeptide-based therapeutics are known in the art. The compositions may be provided in any convenient form including tablets, capsules, troches, powders, sprays, creams, etc. As such the compositions, in pharmaceutically acceptable dosage units or in bulk, may be incorporated into a wide variety of containers. For example, dosage units may be included in a variety of containers including capsules, pills, etc. The compositions may be advantageously combined and/or used in combination with other therapeutic or prophylactic agents, different from the subject compounds. In many instances, administration in conjunction with the subject compositions enhances the efficacy of such agents, see e.g. Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9^th Ed., 1996, McGraw-Hill.

[0023]In another aspect, the invention provides methods of screening for agents which modulate Robo-Slit interactions. These methods generally involve forming a mixture of a Robo-expressing cell, a Slit polypeptide and a candidate agent, and determining the effect of the agent on the amount of Robo expressed by the cell. The methods are amenable to automated, cost-effective high throughput screening of chemical libraries for lead compounds. Identified reagents find use in the pharmaceutical industries for animal and human trials; for example, the reagents may be derivatized and rescreened in in vitro and in vivo assays to optimize activity and minimize toxicity for pharmaceutical development. Cell and animal based neural guidance/repulsion assays are described in detail in the experimental section below.

[0024]The amino acid sequences of the disclosed vertebrate Slit polypeptides are used to back-translate Slit polypeptide-encoding nucleic acids optimized for selected expression systems (Holler et al. (1993) Gene 136, 323-328; Martin et al. (1995) Gene 154, 150-166) or used to generate degenerate oligonucleotide primers and probes for use in the isolation of natural Slit-encoding nucleic acid sequences ("GCG" software, Genetics Computer Group, Inc, Madison Wis.). Slit-encoding nucleic acids used in Slit-expression vectors and incorporated into recombinant host cells, e.g. for expression and screening, transgenic animals, e.g. for functional studies such as the efficacy of candidate drugs for disease associated with Slit-modulated cell function, etc.

[0025]The invention also provides nucleic acid hybridization probes and replication/amplification primers having a vertebrate Slit cDNA specific sequence comprising a fragment of a disclosed vertebrate cDNA sequence, and sufficient to effect specific hybridization thereto. Such primers or probes are at least 12, preferably at least 24, more preferably at least 36 and most preferably at least 96 nucleotides in length. Demonstrating specific hybridization generally requires stringent conditions, for example, hybridizing in a buffer comprising 30% formamide in 5×SSPE (0.18 M NaCl, 0.01 M NaPO₄, pH7.7, 0.001 M EDTA) buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE; preferably hybridizing in a buffer comprising 50% formamide in 5×SSPE buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE buffer at 42° C. Slit nucleic acids can also be distinguished using alignment algorithms, such as BLASTX (Altschul et al. (1990) Basic Local Alignment Search Tool, J Mol Biol 215, 403-410). In addition, the invention provides nucleic acids having a sequence about 60-70%, preferably about 70-80%, more preferably about 80-90%, more preferably about 90-95%, most preferably about 95-99% similar to a vertebrate Slit sequence disclosed herein as determined by Best Fit analysis using default settings and is other than a natural drosophila Slit sequence, preferably other than a natural invertebrate Slit sequence. In a particular embodiment, the Slit polynucleotide fragments comprise species specific fragments; such fragments are readily discerned from alignments of the disclosed sequences.

[0026]The subject nucleic acids are of synthetic/non-natural sequences and/or are recombinant, meaning they comprise a non-natural sequence or a natural sequence joined to nucleotide(s) other than that which it is joined to on a natural chromosome. The subject recombinant nucleic acids comprising the nucleotide sequence of disclosed vertebrate Slit nucleic acids, or fragments thereof, contain such sequence or fragment at a terminus, immediately flanked by (i.e. contiguous with) a sequence other than that which it is joined to on a natural chromosome, or flanked by a native flanking region fewer than 10 kb, preferably fewer than 2 kb, more preferably fewer than 500 bp, which is at a terminus or is immediately flanked by a sequence other than that which it is joined to on a natural chromosome. While the nucleic acids are usually RNA or DNA, it is often advantageous to use nucleic acids comprising other bases or nucleotide analogs to provide modified stability, etc.

[0027]The subject nucleic acids find a wide variety of applications including use as translatable transcripts, hybridization probes, PCR primers, diagnostic nucleic acids, etc.; use in detecting the presence of Slit genes and gene transcripts and in detecting or amplifying nucleic acids encoding additional Slit homologs and structural analogs. In diagnosis, Slit hybridization probes find use in identifying wild-type and mutant Slit alleles in clinical and laboratory samples. Mutant alleles are used to generate allele-specific oligonucleotide (ASO) probes for high-throughput clinical diagnoses. In therapy, therapeutic Slit nucleic acids are used to modulate cellular expression or intracellular concentration or availability of active Slit. Exemplary human Slit-1 probes and primers are shown in Table 5 and Table 6.

TABLE-US-00003 TABLE 5 Hybridization Probes for Regions of Human Slit-1. Hybridization probe for first SEQ ID NO: 01, nucleotides 82-828 leucine rich repeat region Hybridization probe for second SEQ ID NO: 01, nucleotides 829-1503 leucine rich repeat region Hybridization probe for third SEQ ID NO: 01, nucleotides 1504-2166 leucine rich repeat region Hybridization probe for fourth SEQ ID NO: 01, nucleotides 2167-2751 leucine rich repeat region Hybridization probe for EGF SEQ ID NO: 01, nucleotides 2752-3327 repeats one to five Hybridization probe for the SEQ ID NO: 01, nucleotides 3328-3461 sixth EGF repeat and preceding spacer region Hybridization probe for the 99aa SEQ ID NO: 01, nucleotides 3462-3987 spacer/G-loop region Hybridization probe for EGF SEQ ID NO: 01, nucleotides 3988-4341 repeats seven to nine Hybridization probe for the SEQ ID NO: 01, nucleotides 4342-4575 cysteine knot region

TABLE-US-00004 TABLE 6 PCR Primers for regions of Human Slit. PCR Primers for first leucine Forward: SEQ ID NO: 01, nucleotides 82-111 rich repeat region Reverse: reverse complement of SEQ ID NO: 01, nucleotides 799-828 PCR Primers for second Forward: SEQ ID NO: 01, nucleotides 829-858 leucine rich repeat region Reverse: reverse complement of SEQ ID NO: 01, nucleotides 1474-1503 PCR Primers for third leucine Forward: SEQ ID NO: 01, nucleotides 1504-1533 rich repeat region Reverse: reverse complement of SEQ ID NO: 01, nucleotides 2137-2166 PCR Primers for fourth Forward: SEQ ID NO: 01, nucleotides 2167-2196 leucine rich repeat region Reverse: reverse complement of SEQ ID NO: 01, nucleotides 2722-2751 PCR Primers for EGF repeats Forward: SEQ ID NO: 01, nucleotides 2752-2781 one to five Reverse: reverse complement of SEQ ID NO: 01, nucleotides 3298-3327 PCR Primers for the sixth Forward: SEQ ID NO: 01, nucleotides 3328-3357 EGF repeat and preceding Reverse: reverse complement of SEQ ID NO: 01, spacer region nucleotides 3432-3461 PCR Primers for the 99aa Forward: SEQ I: 01, nucleotides 3462-3491 spacer/G-loop region Reverse: reverse complement of SEQ ID NO: 01, nucleotides 3958-3987 PCR Primers for EGF repeats Forward: SEQ ID NO: 01, nucleotides 3988-4017 seven to nine Reverse: reverse complement of SEQ ID NO: 01, nucleotides 4312-4341 PCR Primers for the cysteine Forward: SEQ ID NO: 01, nucleotides 4342-4371 knot region Reverse: reverse complement of SEQ ID NO: 01, nucleotides 4546-4575

[0028]Leucine rich repeats (LRRs) are predicted by comparison with known proteins and by the presence of a leucine rich core sequence. In slit proteins, the LRRs are flanked by conserved sequences referred to as the amino- and carboxy-flanking regions. These flanking regions are found in other known proteins, but only in a few instances are both the amino- and carboxy-flank regions present in a single protein. The so called "99aa spacer" is actually ˜200 amino acids in the Drosophila protein and 174 amino acids in Human Slit-1. This region shows homology to the G-loops of laminin A chains.

[0029]Cysteine knots are dimerisation domains defined by the presence of six cysteine residues between which disulphide bridges form. The only absolutely conserved residues are the six cysteines, and spacing between them is highly variable, apart from between cysteines 2 and 3, and 5 and 6. The glycine between cysteines 2 and 3 is only present in a subset of cysteine knots. Drosophila slit and Human slit-1 both have an extra cysteine after cysteines 5 and 6: this may serve as an intermolecular bond. Human Slit-1 gene displays the overall structure of the Drosophila gene, and amino acid conservation is found along the entire length of the protein (48% homology at the amino acid sequence excluding the signal sequence; see below). The Human gene has an extra LRR between LRR2 and LRR3 of the first set of LRRs; in the third set, the Human gene has an extra LRR between LRR3 and LRR4. The Human gene has two extra EGF repeats, on either side of the seventh EGF repeat in Drosophila slit.

Isolation of Human Slit-1

[0030]Searching of the EST database revealed an EST, ab16g10.r1, with homology to the 99aa spacer region of Drosophila slit. This EST was used to probe a Human fetal brain library (Stratagene), and clones for Human slit-1 were isolated.

TABLE-US-00005 Features of Human Slit Predicted Protein Signal sequence SEQ ID NO: 02, residues 7-24 First amino-flanking sequence SEQ ID NO: 02, residues 28-59 First set of Leucine Rich SEQ ID NO: 02, residues 60-179 Repeats (6 repeats) First carboxy-flanking sequence SEQ ID NO: 02, residues 180-276 Second amino-flanking sequence SEQ ID NO: 02, residues 277-308 Second set of Leucine Rich SEQ ID NO: 02, residues 309-434 Repeats (5 repeats) Second carboxy-flanking sequence SEQ ID NO: 02, residues 435-501 Third amino-flanking sequence SEQ ID NO: 02, residues 502-533 Third set of Leucine Rich SEQ ID NO: 02, residues 534-560 Repeats (5 repeats) Third carboxy-flanking sequence SEQ ID NO: 02, residues 661-722 Fourth amino-flanking sequence SEQ ID NO: 02, residues 723-754 Fourth set of Leucine Rich SEQ ID NO: 02, residues 755-855 Repeats (4 repeats) Fourth carboxy-flanking sequence SEQ ID NO: 02, residues 856-917 First EGF repeat SEQ ID NO: 02, residues 918-952 Second EGF repeat SEQ ID NO: 02, residues 953-993 Third EGF repeat SEQ ID NO: 02, residues 994-1031 Fourth EGF repeat SEQ ID NO: 02, residues 1032-1071 Fifth EGF repeat SEQ ID NO: 02, residues 1072-1109 Spacer SEQ ID NO: 02, residues 1110-1116 Sixth EGF repeat SEQ ID NO: 02, residues 1117-1153 "99aa spacer" SEQ ID NO: 02, residues 1155-1329 Seventh EGF repeat SEQ ID NO: 02, residues 1330-1366 Eighth EGF repeat SEQ ID NO: 02, residues 1367-1404 Ninth EGF repeat SEQ ID NO: 02, residues 1405-1447 Cysteine knot motif SEQ ID NO: 02, residues 1448-1525

TABLE-US-00006 Amino acid identity between Drosophila and Human Slit-1 First amino-flanking sequence 53% First set of Leucine Rich Repeats 52% (54%, 67%, NA, 38%, 54%, 50%) First carboxy-flanking sequence 42% Second amino-flanking sequence 50% Second set of Leucine Rich Repeats 60% (54%, 58%, 67%, 71%, 50%) Second carboxy-flanking sequence 62% Third amino-flanking sequence 56% Third set of Leucine Rich Repeats 49% (46%, 46%, 42%, NA, 58%) Third carboxy-flanking sequence 36% Fourth amino-flanking sequence 53% Fourth set of Leucine Rich Repeats 48% (25%, 58%, 46%, 63%) Fourth carboxy-flanking sequence 63% First EGF repeat 34% Second EGF repeat 46% Third EGF repeat 46% Fourth EGF repeat 35% Fifth EGF repeat 47% Spacer 22% Sixth EGF repeat 40% "99aa spacer" 38% Seventh EGF repeat 11%/NA Eighth EGF repeat 44% Nineth EGF repeat 29%/NA Cysteine knot motif 34% NA: not applicable due to absence of homologous repeat. Figures for individual LLRs are shown in brackets.

[0031]The following examplary assay is offered by way of illustration and not by way of limitation:

EXAMPLES

Protocol for Ligand Screening of Transfected COS Cells

I. Prepare the Ligand

[0032]Expression Construct: cDNAs encoding targeted Slit polypeptides are tagged with the Fc portion of human IgG and subcloned into a 293 expression vector (pCEP4: In Vitrogen).

[0033]Transfection: 293 EBNA cells are transfected (CaPO₄ method) with the Slit expression constructs. After 24 h recovery, transfected cells are selected with G418 (geneticin, 250 ug/ml, Gibco) and hygromycin (200 ug/ml). Once the selection process is complete, cells are maintained in Dulbecco's Modified Eagles medium (DME)/10% FCS under selection.

[0034]Preparation of Conditioned Medium: Serum-containing media is replaced with Optimem with glutamax-1 (Gibco) and 300 ng/ml heparin (Sigma), and the cells are conditioned for 3 days. The media is collected and spun at 3,000×g for 10 minutes. The supernatant is filtered (0.45 um) and stored with 0. 1% azide at 4° C. for no more than 2 weeks.

II. Prepare Truncated Receptor (Positive Control)

[0035]Expression Construct: cDNA encoding a corresponding Robo C-terminal deletion mutant comprising the extracellular domain (truncated immediately N-terminal to the transmembrane region) is subcloned into a 293 expression vector (pCEP4: In Vitrogen).

[0036]Transfection: 293 EBNA cells are transfected (CaPO₄ method) with the receptor mutant expression construct. After 24 h recovery, transfected cells are selected with G418 (geneticin, 250 ug/ml, Gibco) and hygromycin (200 ug/ml). Once the selection process is complete, cells are maintained in Dulbecco's Modified Eagles medium (DME)/10% FCS under selection.

[0037]Preparation of Conditioned Medium: Serum-containing media is replaced with Optimem with glutamax-1 (Gibco) and 300 ng/ml heparin (Sigma), and the cells are conditioned for 3 days. The media is collected and spun at 3,000×g for 10 minutes. The supernatant is filtered (0.45 um) and stored with 0.1% azide at 4° C. for no more than 2 weeks.

II. Transfect COS Cells

[0038]Seed COS cells (250,000) on 35 mm dishes in 2 ml DME/10% FCS.

[0039]18-24 h later, dilute 1 ug of Robo-encoding DNA (cDNA cloned into pMT21 expression vector) into 200 ul serum-free media and add 6 ul of Lipofectamine (Gibco). Incubate this solution at room temperature for 15-45 min.

[0040]Wash the cells 2× with PBS. Add 800 ul serum-free media to the tube containing the lipid-DNA complexes. Overlay this solution onto the washed cells.

[0041]Incubate for 6 h. Stop the reaction by adding 1 ml DMA/20% FCS. Refeed cells. Assay cells 12 hr later.

III. Ligand Binding Assay

[0042]Wash plates of transfected COS cells IX with cold PBS (plus Ca/Mg)/1% goat serum. Add 1 ml conditioned media neat and incubate 90 min at room temp.

[0043]Wash plates 3× with PBS (plus Ca/Mg). On the 4th wash, add 1 ml 50% methanol to 1 ml PBS. Then add 1 ml methanol. Evacuate and add 1 ml methanol.

[0044]Wash 1× with PBS. Wash 1× PBS/1% goat serum.

[0045]Add secondary antibody (1-to-2,000 anti-human Fc conjugated to alkaline phosphatase (Jackson Lab)) in PBS/1% goat serum. Incubate 30-40 min room temp.

[0046]Wash 3× with PBS. Wash 1× alkaline phosphatase buffer (100 mM Tris-Cl, pH 9.5, 100 mM NaCl, 5 mM MgCl₂). Prepare alkaline phosphatase reagents: 4.5 ul/ml NBT and 3.5 ul/ml BCIP (Gibco) in alkaline phosphatase buffer.

[0047]Incubate 10-30 min, quench with 20 mM EDTA in PBS. Cells that have bound Slit polypeptides are visible by the presence of a dark purple reaction product.

[0048]In parallel incubations, positive controls are provided by titrating Slit binding with serial dilutions of the mutant receptor conditioned medium.

IV. Results: Binding of Slit to Robo

[0049]Cell expressing mammalian Slit polypeptides were shown to bind Robo. No reactivity was observed with control COS cells or with receptor-expressing COS cells in the presence of the secondary antibody but in the absence of the Slit-Fc fusion. Binding was observed to receptor-expression cells using a construct in which a Slit polypeptide is fused directly to alkaline phosphatase, for which a secondary antibody is not required. Receptor deletion mutants titrate the Slit-Robo binding, serving as a positive control for inhibition assays.

Protocol for High Throughput Robo-Slit Binding Assay

A. Reagents:

[0050]Neutralite Avidin: 20 μg/ml in PBS.

[0051]Blocking buffer: 5% BSA, 0.5% Tween 20 in PBS; 1 hour at room temperature.

[0052]Assay Buffer: 100 mM KCl, 20 mM HEPES pH 7.6, 1 mM MgCl₂, 1% glycerol, 0.5% NP-40, 50 mM β-mercaptoethanol, 1 mg/ml BSA, cocktail of protease inhibitors.

[0053]³³P Robo polypeptide 10× stock: 10^-8-10^-6 M "cold" Robo polypeptide specific Robo domain supplemented with 200,000-250,000 cpm of labeled Robo (Beckman counter). Place in the 4° C. microfridge during screening.

[0054]Protease inhibitor cocktail (1000×): 10 mg Trypsin Inhibitor (BMB # 109894), 10 mg Aprotinin (BMB # 236624), 25 mg Benzamidine (Sigma # B-6506), 25 mg Leupeptin (BMB # 1017128), 10 mg APMSF (BMB #917575), and 2 mM NaVO₃ (Sigma #S-6508) in 10 ml of PBS.

[0055]Slit: 10^-7-10^-5 M biotinylated Slit in PBS.

B. Preparation of Assay Plates

[0056]Coat with 120 μl of stock N-Avidin per well overnight at 4° C.

[0057]Wash 2 times with 200 μl PBS.

[0058]Block with 150 μl of blocking buffer.

[0059]Wash 2 times with 200 μl PBS.

C. Assay

[0060]Add 40 μl assay buffer/well.

[0061]Add 10 μl compound or extract.

[0062]Add 10 μl ³³P-Robo (20-25,000 cpm/0.1-10 pmoles/well=10^-9-10^-7 M final conc).

[0063]Shake at 25° C. for 15 minutes.

[0064]Incubate additional 45 minutes at 25° C.

[0065]Add 40 μM biotinylated Slit (0.1-10 pmoles/40 ul in assay buffer)

[0066]Incubate 1 hour at room temperature.

[0067]Stop the reaction by washing 4 times with 200 μM PBS.

[0068]Add 150 μM scintillation cocktail.

[0069]Count in Topcount.

D. Controls for All Assays (Located on Each Plate)

[0070]a. Non-specific binding

[0071]b. Soluble (non-biotinylated Slit) at 80% inhibition. [0072]All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Sequence CWU 1

2014758DNAhumanCDS(1)..(4575) 1atg cgc ggc gtt ggc tgg cag atg ctg tcc ctg tcg ctg ggg tta gtg 48Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val1 5 10 15ctg gcg atc ctg aac aag gtg gca ccg cag gcg tgc ccg gcg cag tgc 96Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys20 25 30tct tgc tcg ggc agc aca gtg gac tgt cac ggg ctg gcg ctg cgc agc 144Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser35 40 45gtg ccc agg aat atc ccc cgc aac acc gag aga ctg gat tta aat gga 192Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly50 55 60aat aac atc aca aga att acg aag aca gat ttt gct ggt ctt aga cat 240Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His65 70 75 80cta aga gtt ctt cag ctt atg gag aat aag att agc acc att gaa aga 288Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg85 90 95gga gca ttc cag gat ctt aaa gaa cta gag aga ctg cgt tta aac aga 336Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg100 105 110aat cac ctt cag ctg ttt cct gag ttg ctg ttt ctt ggg act gcg aag 384Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys115 120 125cta tac agg ctt gat ctc agt gaa aac caa att cag gca atc cca agg 432Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg130 135 140aaa gct ttc cgt ggg gca gtt gac ata aaa aat ttg caa ctg gat tac 480Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr145 150 155 160aac cag atc agc tgt att gaa gat ggg gca ttc agg gct ctc cgg gac 528Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp165 170 175ctg gaa gtg ctc act ctc aac aat aac aac att act aga ctt tct gtg 576Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val180 185 190gca agt ttc aac cat atg cct aaa ctt agg act ttt cga ctg cat tca 624Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser195 200 205aac aac ctg tat tgt gac tgc cac ctg gcc tgg ctc tcc gac tgg ctt 672Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu210 215 220cgc aaa agg cct cgg gtt ggt ctg tac act cag tgt atg ggc ccc tcc 720Arg Lys Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser225 230 235 240cac ctg aga ggc cat aat gta gcc gag gtt caa aaa cga gaa ttt gtc 768His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val245 250 255tgc agt gat gag gaa gaa ggt cac cag tca ttt atg gct cct tct tgt 816Cys Ser Asp Glu Glu Glu Gly His Gln Ser Phe Met Ala Pro Ser Cys260 265 270agt gtt ttg cac tgc cct gcc gcc tgt acc tgt agc aac aat atc gta 864Ser Val Leu His Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val275 280 285gac tgt cgt ggg aaa ggt ctc act gag atc ccc aca aat ctt cca gag 912Asp Cys Arg Gly Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu290 295 300acc atc aca gaa ata cgt ttg gaa cag aac aca atc aaa gtc atc cct 960Thr Ile Thr Glu Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro305 310 315 320cct gga gct ttc tca cca tat aaa aag ctt aga cga att gac ctg agc 1008Pro Gly Ala Phe Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser325 330 335aat aat cag atc tct gaa ctt gca cca gat gct ttc caa gga cta cgc 1056Asn Asn Gln Ile Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg340 345 350tct ctg aat tca ctt gtc ctc tat gga aat aaa atc aca gaa ctc ccc 1104Ser Leu Asn Ser Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro355 360 365aaa agt tta ttt gaa gga ctg ttt tcc tta cag ctc cta tta ttg aat 1152Lys Ser Leu Phe Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn370 375 380gcc aac aag ata aac tgc ctt cgg gta gat gct ttt cag gat ctc cac 1200Ala Asn Lys Ile Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His385 390 395 400aac ttg aac ctt ctc tcc cta tat gac aac aag ctt cag acc atc gcc 1248Asn Leu Asn Leu Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala405 410 415aag ggg acc ttt tca cct ctt cgg gcc att caa act atg cat ttg gcc 1296Lys Gly Thr Phe Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala420 425 430cag aac ccc ttt att tgt gac tgc cat ctc aag tgg cta gcg gat tat 1344Gln Asn Pro Phe Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr435 440 445ctc cat acc aac ccg att gag acc agt ggt gcc cgt tgc acc agc ccc 1392Leu His Thr Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro450 455 460cgc cgc ctg gca aac aaa aga att gga cag atc aaa agc aag aaa ttc 1440Arg Arg Leu Ala Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe465 470 475 480cgt tgt tca ggt aca gaa gat tat cga tca aaa tta agt gga gac tgc 1488Arg Cys Ser Gly Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys485 490 495ttt gcg gat ctg gct tgc cct gaa aag tgt cgc tgt gaa gga acc aca 1536Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr500 505 510gta gat tgc tct aat caa aag ctc aac aaa atc ccg gag cac att ccc 1584Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro515 520 525cag tac act gca gag ttg cgt ctc aat aat aat gaa ttt acc gtg ttg 1632Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu530 535 540gaa gcc aca gga atc ttt aag aaa ctt cct caa tta cgt aaa ata aac 1680Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn545 550 555 560ttt agc aac aat aag atc aca gat att gag gag gga gca ttt gaa gga 1728Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly565 570 575gca tct ggt gta aat gaa ata ctt ctt acg agt aat cgt ttg gaa aat 1776Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn580 585 590gtg cag cat aag atg ttc aag gga ttg gaa agc ctc aaa act ttg atg 1824Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met595 600 605ttg aga agc aat cga ata acc tgt gtg ggg aat gac agt ttc ata gga 1872Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly610 615 620ctc agt tct gtg cgt ttg ctt tct ttg tat gat aat caa att act aca 1920Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr625 630 635 640gtt gca cca ggg gca ttt gat act ctc cat tct tta tct act cta aac 1968Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn645 650 655ctc ttg gcc aat cct ttt aac tgt aac tgc tac ctg gct tgg ttg gga 2016Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly660 665 670gag tgg ctg aga aag aag aga att gtc acg gga aat cct aga tgt caa 2064Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln675 680 685aaa cca tac ttc ctg aaa gaa ata ccc atc cag gat gtg gcc att cag 2112Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln690 695 700gac ttc act tgt gat gac gga aat gat gac aat agt tgc tcc cca ctt 2160Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu705 710 715 720tct cgc tgt cct act gaa tgt act tgc ttg gat aca gtc gtc cga tgt 2208Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys725 730 735agc aac aag ggt ttg aag gtc ttg ccg aaa ggt att cca aga gat gtc 2256Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val740 745 750aca gag ttg tat ctg gat gga aac caa ttt aca ctg gtt ccc aag gaa 2304Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu755 760 765ctc tcc aac tac aaa cat tta aca ctt ata gac tta agt aac aac aga 2352Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg770 775 780ata agc acg ctt tct aat cag agc ttc agc aac atg acc cag ctc ctc 2400Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu785 790 795 800acc tta att ctt agt tac aac cgt ctg aga tgt att cct cct cgc acc 2448Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr805 810 815ttt gat gga tta aag tct ctt cga tta ctt tct cta cat gga aat gac 2496Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp820 825 830att tct gtt gtg cct gaa ggt gct ttc aat gat ctt tct gca tta tca 2544Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser835 840 845cat cta gca att gga gcc aac cct ctt tac tgt gat tgt aac atg cag 2592His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln850 855 860tgg tta tcc gac tgg gtg aag tcg gaa tat aag gag cct gga att gct 2640Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala865 870 875 880cgt tgt gct ggt cct gga gaa atg gca gat aaa ctt tta ctc aca act 2688Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr885 890 895ccc tcc aaa aaa ttt acc tgt caa ggt cct gtg gat gtc aat att cta 2736Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu900 905 910gct aag tgt aac ccc tgc cta tca aat ccg tgt aaa aat gat ggc aca 2784Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr915 920 925tgt aat agt gat cca gtt gac ttt tac cga tgc acc tgt cca tat ggt 2832Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly930 935 940ttc aag ggg cag gac tgt gat gtc cca att cat gcc tgc atc agt aac 2880Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn945 950 955 960cca tgt aaa cat gga gga act tgc cac tta aag gaa gga gaa gaa gat 2928Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp965 970 975gga ttc tgg tgt att tgt gct gat gga ttt gaa gga gaa aat tgt gaa 2976Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu980 985 990gtc aac gtt gat gat tgt gaa gat aat gac tgt gaa aat aat tct aca 3024Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr995 1000 1005tgt gtc gat ggc att aat aac tac aca tgc ctt tgc cca cct gag tat 3072Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr1010 1015 1020aca ggt gag ttg tgt gag gag aag ctg gac ttc tgt gcc cag gac ctg 3120Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu1025 1030 1035 1040aac ccc tgc cag cac gat tca aag tgc atc cta act cca aag gga ttc 3168Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe1045 1050 1055aaa tgt gac tgc aca cca ggg tac gta ggt gaa cac tgc gac atc gat 3216Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp1060 1065 1070ttt gac gac tgc caa gac aac aag tgt aaa aac gga gcc cac tgc aca 3264Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr1075 1080 1085gat gca gtg aac ggc tat acg tgc ata tgc ccc gaa ggt tac agt ggc 3312Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly1090 1095 1100ttg ttc tgt gag ttt tct cca ccc atg gtc ctc cct cgt acc agc ccc 3360Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro1105 1110 1115 1120tgt gat aat ttt gat tgt cag aat gga gct cag tgt atc gtc aga ata 3408Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile1125 1130 1135aat gag cca ata tgt cag tgt ttg cct ggc tat cag gga gaa aag tgt 3456Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys1140 1145 1150gaa aaa ttg gtt agt gtg aat ttt ata aac aaa gag tct tat ctt cag 3504Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln1155 1160 1165att cct tca gcc aag gtt cgg cct cag acg aac ata aca ctt cag att 3552Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile1170 1175 1180gcc aca gat gaa gac agc gga atc ctc ctg tat aag ggt gac aaa gac 3600Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp1185 1190 1195 1200cat atc gcg gta gaa ctc tat cgg ggg cgt gtt cgt gcc agc tat gac 3648His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg Ala Ser Tyr Asp1205 1210 1215acc ggc tct cat cca gct tct gcc att tac agt gtg gag aca atc aat 3696Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn1220 1225 1230gat gga aac ttc cac att gtg gaa cta ctt gcc ttg gat cag agt ctc 3744Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu1235 1240 1245tct ttg tcc gtg gat ggt ggg aac ccc aaa atc atc act aac ttg tca 3792Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser1250 1255 1260aag cag tcc act ctg aat ttt gac tct cca ctc tat gta gga ggc atg 3840Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met1265 1270 1275 1280cca ggg aag agt aac gtg gca tct ctg cgc cag gcc cct ggg cag aac 3888Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn1285 1290 1295gga acc agc ttc cac ggc tgc atc cgg aac ctt tac atc aac agt gag 3936Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu1300 1305 1310ctg cag gac ttc cag aag gtg ccg atg caa aca ggc att ttg cct ggc 3984Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly1315 1320 1325tgt gag cca tgc cac aag aag gtg tgt gcc cat ggc aca tgc cag ccc 4032Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro1330 1335 1340agc agc cag gca ggc ttc acc tgc gag tgc cag gaa gga tgg atg ggg 4080Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly1345 1350 1355 1360ccc ctc tgt gac caa cgg acc aat gac cct tgc ctt gga aat aaa tgc 4128Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys1365 1370 1375gta cat ggc acc tgc ttg ccc atc aat gcg ttc tcc tac agc tgt aag 4176Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys1380 1385 1390tgc ttg gag ggc cat gga ggt gtc ctc tgt gat gaa gag gag gat ctg 4224Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu1395 1400 1405ttt aac cca tgc cag gcg atc aag tgc aag cat ggg aag tgc agg ctt 4272Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu1410 1415 1420tca ggt ctg ggg cag ccc tac tgt gaa tgc agc agt gga tac acg ggg 4320Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly1425 1430 1435 1440gac agc tgt gat cga gaa atc tct tgt cga ggg gaa agg ata aga gat 4368Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp1445 1450 1455tat tac caa aag cag cag ggc tat gct gct tgc caa aca acc aag aag 4416Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys1460 1465 1470gtg tcc cga tta gag tgc aga ggt ggg tgt gca gga ggg cag tgc tgt 4464Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys1475 1480 1485gga ccg ctg agg agc aag cgg cgg aaa tac tct ttc gaa tgc act gac 4512Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp1490 1495 1500ggc tcc tcc ttt gtg gac gag gtt gag aaa gtg gtg aag tgc ggc tgt 4560Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys1505 1510 1515 1520acg agg tgt gtg tcc taaacacact cccggcagct ctgtctttgg aaaaggttgt 4615Thr Arg Cys Val Ser1525atacttcttg accatgtggg actaatgaat gcttcatagt ggaaatattt gaaatatatt 4675gtaaaataca gaacagactt atttttatta tgagaataaa gacttttttt ctgcatttgg 4735aaaaaaaaaa aaaaaaaact cga 475821525PRThuman 2Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys20 25 30Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser35 40 45Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly50 55 60Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His65 70 75

80Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg85 90 95Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg100 105 110Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys115 120 125Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg130 135 140Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr145 150 155 160Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp165 170 175Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val180 185 190Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser195 200 205Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu210 215 220Arg Lys Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser225 230 235 240His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val245 250 255Cys Ser Asp Glu Glu Glu Gly His Gln Ser Phe Met Ala Pro Ser Cys260 265 270Ser Val Leu His Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val275 280 285Asp Cys Arg Gly Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu290 295 300Thr Ile Thr Glu Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro305 310 315 320Pro Gly Ala Phe Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser325 330 335Asn Asn Gln Ile Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg340 345 350Ser Leu Asn Ser Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro355 360 365Lys Ser Leu Phe Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn370 375 380Ala Asn Lys Ile Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His385 390 395 400Asn Leu Asn Leu Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala405 410 415Lys Gly Thr Phe Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala420 425 430Gln Asn Pro Phe Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr435 440 445Leu His Thr Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro450 455 460Arg Arg Leu Ala Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe465 470 475 480Arg Cys Ser Gly Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys485 490 495Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr500 505 510Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro515 520 525Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu530 535 540Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn545 550 555 560Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly565 570 575Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn580 585 590Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met595 600 605Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly610 615 620Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr625 630 635 640Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn645 650 655Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly660 665 670Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln675 680 685Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln690 695 700Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu705 710 715 720Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys725 730 735Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val740 745 750Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu755 760 765Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg770 775 780Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu785 790 795 800Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr805 810 815Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp820 825 830Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser835 840 845His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln850 855 860Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala865 870 875 880Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr885 890 895Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu900 905 910Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr915 920 925Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly930 935 940Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn945 950 955 960Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp965 970 975Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu980 985 990Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr995 1000 1005Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr1010 1015 1020Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu1025 1030 1035 1040Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe1045 1050 1055Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp1060 1065 1070Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr1075 1080 1085Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly1090 1095 1100Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro1105 1110 1115 1120Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile1125 1130 1135Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys1140 1145 1150Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln1155 1160 1165Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile1170 1175 1180Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp1185 1190 1195 1200His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg Ala Ser Tyr Asp1205 1210 1215Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn1220 1225 1230Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu1235 1240 1245Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser1250 1255 1260Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met1265 1270 1275 1280Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn1285 1290 1295Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu1300 1305 1310Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly1315 1320 1325Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro1330 1335 1340Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly1345 1350 1355 1360Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys1365 1370 1375Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys1380 1385 1390Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu1395 1400 1405Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu1410 1415 1420Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly1425 1430 1435 1440Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp1445 1450 1455Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys1460 1465 1470Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys1475 1480 1485Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp1490 1495 1500Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys1505 1510 1515 1520Thr Arg Cys Val Ser15253105PRThuman 3Ser Pro Cys Thr Cys Ser Asn Asn Ile Val Asp Cys Arg Gly Lys Gly 1 5 10 15Leu Met Glu Ile Pro Ala Asn Leu Pro Glu Gly Ile Val Glu Ile Arg20 25 30Leu Glu Gln Asn Ser Ile Lys Ala Ile Pro Ala Gly Ala Phe Thr Gln35 40 45Tyr Lys Lys Leu Lys Arg Ile Asp Ile Ser Lys Asn Gln Ile Ser Asp50 55 60Ile Ala Pro Asp Ala Phe Gln Gly Leu Lys Ser Leu Thr Ser Leu Val65 70 75 80Leu Tyr Gly Asn Lys Ile Thr Glu Ile Ala Lys Gly Leu Phe Asp Gly85 90 95Leu Val Ser Leu Gln Leu Leu Leu Leu100 1054138PRThuman 4Glu Gly Ala Phe Asn Gly Ala Ala Ser Val Gln Glu Leu Met Leu Thr1 5 10 15Gly Asn Gln Leu Glu Thr Val His Gly Arg Gly Phe Arg Gly Gly Leu20 25 30Ser Gly Leu Lys Thr Leu Met Leu Arg Ser Asn Leu Ile Gly Cys Val35 40 45Ser Asn Asp Thr Phe Ala Gly Leu Ser Ser Val Arg Leu Leu Ser Leu50 55 60Tyr Asp Asn Arg Ile Thr Thr Ile Thr Pro Gly Ala Phe Thr Thr Leu65 70 75 80Val Ser Leu Ser Thr Ile Asn Leu Leu Ser Asn Pro Phe Asn Cys Asn85 90 95Cys His Leu Gly Ala Gly Leu Gly Lys Trp Leu Arg Lys Arg Arg Ile100 105 110Val Ser Gly Asn Pro Arg Cys Gln Lys Pro Phe Phe Leu Lys Glu Ile115 120 125Pro Ile Gln Gly Val Gly His Pro Gly Ile130 1355160PRThumanmisc_feature(121)..(150)note="Xaa signifies gap in sequence" 5Trp Pro Arg Cys Glu Cys Met Pro Gly Tyr Ala Gly Asp Asn Cys Ser1 5 10 15Glu Asn Gln Asp Asp Cys Arg Asp His Arg Cys Gln Asn Gly Ala Gln20 25 30Cys Met Asp Glu Val Asn Ser Tyr Ser Cys Leu Cys Ala Glu Gly Tyr35 40 45Ser Gly Gln Leu Cys Glu Ile Pro Pro His Leu Pro Ala Pro Lys Ser50 55 60Pro Cys Glu Gly Thr Glu Cys Gln Asn Gly Ala Asn Cys Val Asp Gln65 70 75 80Gly Asn Arg Pro Val Cys Gln Cys Leu Pro Gly Phe Gly Gly Pro Glu85 90 95Cys Glu Lys Leu Leu Ser Val Asn Phe Val Asp Arg Asp Thr Tyr Leu100 105 110Gln Phe Thr Asp Leu Gln Asn Trp Xaa Arg Xaa Asn Ile Thr Leu Gln115 120 125Val Phe Thr Ala Glu Asp Asn Gly Ile Leu Leu Tyr Asn Gly Gly Asn130 135 140Asp His Ile Ala Val Xaa Leu Tyr Xaa Gly His Val Arg Phe Ser Tyr145 150 155 1606103PRThuman 6Gln Cys His Ile Ser Asp Gln Gly Glu Pro Tyr Cys Leu Cys Gln Pro1 5 10 15Gly Phe Ser Gly Glu His Cys Gln Gln Glu Asn Pro Cys Leu Gly Gln20 25 30Val Val Arg Glu Val Ile Arg Arg Gln Lys Gly Tyr Ala Ser Cys Ala35 40 45Thr Ala Ser Lys Val Pro Ile Met Glu Cys Arg Gly Gly Cys Gly Pro50 55 60Gln Cys Cys Gln Pro Thr Arg Ser Lys Arg Arg Lys Tyr Val Phe Gln65 70 75 80Cys Thr Asp Gly Ser Ser Phe Val Glu Glu Val Glu Arg His Leu Glu85 90 95Cys Gly Cys Leu Ala Cys Ser10071480PRTDrosophila melanogaster 7Met Ala Ala Pro Ser Arg Thr Thr Leu Met Pro Pro Pro Phe Arg Leu1 5 10 15Gln Leu Arg Leu Leu Ile Leu Pro Ile Leu Leu Leu Leu Arg His Asp20 25 30Ala Val His Ala Glu Pro Tyr Ser Gly Gly Phe Gly Ser Ser Ala Val35 40 45Ser Ser Gly Gly Leu Gly Ser Val Gly Ile His Ile Pro Gly Gly Gly50 55 60Val Gly Val Ile Thr Glu Ala Arg Cys Pro Arg Val Cys Ser Cys Thr65 70 75 80Gly Leu Asn Val Asp Cys Ser His Arg Gly Leu Thr Ser Val Pro Arg85 90 95Lys Ile Ser Ala Asp Val Glu Arg Leu Glu Leu Gln Gly Asn Asn Leu100 105 110Thr Val Ile Tyr Glu Thr Asp Phe Gln Arg Leu Thr Lys Leu Arg Met115 120 125Leu Gln Leu Thr Asp Asn Gln Ile His Thr Ile Glu Arg Asn Ser Phe130 135 140Gln Asp Leu Val Ser Leu Glu Arg Leu Asp Ile Ser Asn Asn Val Ile145 150 155 160Thr Thr Val Gly Arg Arg Val Phe Lys Gly Ala Gln Ser Leu Arg Ser165 170 175Leu Gln Leu Asp Asn Asn Gln Ile Thr Cys Leu Asp Glu His Ala Phe180 185 190Lys Gly Leu Val Glu Leu Glu Ile Leu Thr Leu Asn Asn Asn Asn Leu195 200 205Thr Ser Leu Pro His Asn Ile Phe Gly Gly Leu Gly Arg Leu Arg Ala210 215 220Leu Arg Leu Ser Asp Asn Pro Phe Ala Cys Asp Cys His Leu Ser Trp225 230 235 240Leu Ser Arg Phe Leu Arg Ser Ala Thr Arg Leu Ala Pro Tyr Thr Arg245 250 255Cys Gln Ser Pro Ser Gln Leu Lys Gly Gln Asn Val Ala Asp Leu His260 265 270Asp Gln Glu Phe Lys Cys Ser Gly Leu Thr Glu His Ala Pro Met Glu275 280 285Cys Gly Ala Glu Asn Ser Cys Pro His Pro Cys Arg Cys Ala Asp Gly290 295 300Ile Val Asp Cys Arg Glu Lys Ser Leu Thr Ser Val Pro Val Thr Leu305 310 315 320Pro Asp Asp Thr Thr Asp Val Arg Leu Glu Gln Asn Phe Ile Thr Glu325 330 335Leu Pro Pro Lys Ser Phe Ser Ser Phe Arg Arg Leu Arg Arg Ile Asp340 345 350Leu Ser Asn Asn Asn Ile Ser Arg Ile Ala His Asp Ala Leu Ser Gly355 360 365Leu Lys Gln Leu Thr Thr Leu Val Leu Tyr Gly Asn Lys Ile Lys Asp370 375 380Leu Pro Ser Gly Val Phe Lys Gly Leu Gly Ser Leu Arg Leu Leu Leu385 390 395 400Leu Asn Ala Asn Glu Ile Ser Cys Ile Arg Lys Asp Ala Phe Arg Asp405 410 415Leu His Ser Leu Ser Leu Leu Ser Leu Tyr Asp Asn Asn Ile Gln Ser420 425 430Leu Ala Asn Gly Thr Phe Asp Ala Met Lys Ser Met Lys Thr Val His435 440 445Leu Ala Lys Asn Pro Phe Ile Cys Asp Cys Asn Leu Arg Trp Leu Ala450 455 460Asp Tyr Leu His Lys Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Glu465 470 475 480Ser Pro Lys Arg Met His Arg Arg Arg Ile Glu Ser Leu Arg Glu Glu485 490 495Lys Phe Lys Cys Ser Trp Gly Glu Leu Arg Met Lys Leu Ser Gly Glu500 505 510Cys Arg Met Asp Ser Asp Cys Pro Ala Met Cys His Cys Glu Gly Thr515 520 525Thr Val Asp Cys Thr Gly Arg Arg Leu Lys Glu Ile Pro Arg Asp Ile530 535 540Pro Leu His Thr Thr Glu Leu Leu Leu Asn Asp Asn Glu Leu Gly Arg545 550 555 560Ile Ser Ser Asp Gly Leu Phe Gly Arg Leu Pro His Leu Val Lys Leu565 570 575Glu Leu Lys Arg Asn Gln Leu Thr Gly Ile Glu Pro Asn Ala Phe Glu580 585 590Gly Ala Ser His Ile Gln Glu Leu Gln Leu Gly Glu Asn Lys Ile Lys595 600 605Glu Ile Ser Asn Lys Met Phe Leu Gly Leu His Gln Leu Lys Thr Leu610 615 620Asn Leu Tyr Asp Asn Gln Ile Ser Cys Val Met Pro Gly Ser Phe Glu625 630 635 640His Leu Asn Ser Leu Thr Ser Leu Asn Leu Ala Ser Asn Pro Phe Asn645 650 655Cys Asn Cys His Leu Ala Trp Phe Ala Glu Cys Val Arg Lys Lys Ser660 665 670Leu Asn Gly Gly Ala Ala Arg Cys Gly Ala Pro Ser Lys Val Arg

Asp675 680 685Val Gln Ile Lys Asp Leu Pro His Ser Glu Phe Lys Cys Ser Ser Glu690 695 700Asn Ser Glu Gly Cys Leu Gly Asp Gly Tyr Cys Pro Pro Ser Cys Thr705 710 715 720Cys Thr Gly Thr Val Val Ala Cys Ser Arg Asn Gln Leu Lys Glu Ile725 730 735Pro Arg Gly Ile Pro Ala Glu Thr Ser Glu Leu Tyr Leu Glu Ser Asn740 745 750Glu Ile Glu Gln Ile His Tyr Glu Arg Ile Arg His Leu Arg Ser Leu755 760 765Thr Arg Leu Asp Leu Ser Asn Asn Gln Ile Thr Ile Leu Ser Asn Tyr770 775 780Thr Phe Ala Asn Leu Thr Lys Leu Ser Thr Leu Ile Ile Ser Tyr Asn785 790 795 800Lys Leu Gln Cys Leu Gln Arg His Ala Leu Ser Gly Leu Asn Asn Leu805 810 815Arg Val Val Ser Leu His Gly Asn Arg Ile Ser Met Leu Pro Glu Gly820 825 830Ser Phe Glu Asp Leu Lys Ser Leu Thr His Ile Ala Leu Gly Ser Asn835 840 845Pro Leu Tyr Cys Asp Cys Gly Leu Lys Trp Phe Ser Asp Trp Ile Lys850 855 860Leu Asp Tyr Val Glu Pro Gly Ile Ala Arg Cys Ala Glu Pro Glu Gln865 870 875 880Met Lys Asp Lys Leu Ile Leu Ser Thr Pro Ser Ser Ser Phe Val Cys885 890 895Arg Gly Arg Val Arg Asn Asp Ile Leu Ala Lys Cys Asn Ala Cys Phe900 905 910Glu Gln Pro Cys Gln Asn Gln Ala Gln Cys Val Ala Leu Pro Gln Arg915 920 925Glu Tyr Gln Cys Leu Cys Gln Pro Gly Tyr His Gly Lys His Cys Glu930 935 940Phe Met Ile Asp Ala Cys Tyr Gly Asn Pro Cys Arg Asn Asn Ala Thr945 950 955 960Cys Thr Val Leu Glu Glu Gly Arg Phe Ser Cys Gln Cys Ala Pro Gly965 970 975Tyr Thr Gly Ala Arg Cys Glu Thr Asn Ile Asp Asp Cys Leu Gly Glu980 985 990Ile Lys Cys Gln Asn Asn Ala Thr Cys Ile Asp Gly Val Glu Ser Tyr995 1000 1005Lys Cys Glu Cys Gln Pro Gly Phe Ser Gly Glu Phe Cys Asp Thr Lys1010 1015 1020Ile Gln Phe Cys Ser Pro Glu Phe Asn Pro Cys Ala Asn Gly Ala Lys1025 1030 1035 1040Cys Met Asp His Phe Thr His Tyr Ser Cys Asp Cys Gln Ala Gly Phe1045 1050 1055His Gly Thr Asn Cys Thr Asp Asn Ile Asp Asp Cys Gln Asn His Met1060 1065 1070Cys Gln Asn Gly Gly Thr Cys Val Asp Gly Ile Asn Asp Tyr Gln Cys1075 1080 1085Arg Cys Pro Asp Asp Tyr Thr Gly Lys Tyr Cys Glu Gly His Asn Met1090 1095 1100Ile Ser Met Met Tyr Pro Gln Thr Ser Pro Cys Gln Asn His Glu Cys1105 1110 1115 1120Lys His Gly Val Cys Phe Gln Pro Asn Ala Gln Gly Ser Asp Tyr Leu1125 1130 1135Cys Arg Cys His Pro Gly Tyr Thr Gly Lys Trp Cys Glu Tyr Leu Thr1140 1145 1150Ser Ile Ser Phe Val His Asn Asn Ser Phe Val Glu Leu Glu Pro Leu1155 1160 1165Arg Thr Arg Pro Glu Ala Asn Val Thr Ile Val Phe Ser Ser Ala Glu1170 1175 1180Gln Asn Gly Ile Leu Met Tyr Asp Gly Gln Asp Ala His Leu Ala Val1185 1190 1195 1200Glu Leu Phe Asn Gly Arg Ile Arg Val Ser Tyr Asp Val Gly Asn His1205 1210 1215Pro Val Ser Thr Met Tyr Ser Phe Glu Met Val Ala Asp Gly Lys Tyr1220 1225 1230His Ala Val Glu Leu Leu Ala Ile Lys Lys Asn Phe Thr Leu Arg Val1235 1240 1245Asp Arg Gly Leu Ala Arg Ser Ile Ile Asn Glu Gly Ser Asn Asp Tyr1250 1255 1260Leu Lys Leu Thr Thr Pro Met Phe Leu Gly Gly Leu Pro Val Asp Pro1265 1270 1275 1280Ala Gln Gln Ala Tyr Lys Asn Trp Gln Ile Arg Asn Leu Thr Ser Phe1285 1290 1295Lys Gly Cys Met Lys Glu Val Trp Ile Asn His Lys Leu Val Asp Phe1300 1305 1310Gly Asn Ala Gln Arg Gln Gln Lys Ile Thr Pro Gly Cys Ala Leu Leu1315 1320 1325Glu Gly Glu Gln Gln Glu Glu Glu Asp Asp Glu Gln Asp Phe Met Asp1330 1335 1340Glu Thr Pro His Ile Lys Glu Glu Pro Val Asp Pro Cys Leu Glu Asn1345 1350 1355 1360Lys Cys Arg Arg Gly Ser Arg Cys Val Pro Asn Ser Asn Ala Arg Asp1365 1370 1375Gly Tyr Gln Cys Lys Cys Lys His Gly Gln Arg Gly Arg Tyr Cys Asp1380 1385 1390Gln Gly Glu Gly Ser Thr Glu Pro Pro Thr Val Thr Ala Ala Ser Thr1395 1400 1405Cys Arg Lys Glu Gln Val Arg Glu Tyr Tyr Thr Glu Asn Asp Cys Arg1410 1415 1420Ser Arg Gln Pro Leu Lys Tyr Ala Lys Cys Val Gly Gly Cys Gly Asn1425 1430 1435 1440Gln Cys Cys Ala Ala Lys Ile Val Arg Arg Arg Lys Val Arg Met Val1445 1450 1455Cys Ser Asn Asn Arg Lys Tyr Ile Lys Asn Leu Asp Ile Val Arg Lys1460 1465 1470Cys Gly Cys Thr Lys Lys Cys Tyr1475 14808155PRTCaenorhabditis elegansmisc_feature(4)..(152)note="Xaa signifies gap in sequence" 8Arg Asn Pro Xaa Ile Cys Asp Cys Asn Leu Gln Trp Leu Ala Gln Ile1 5 10 15Asn Leu Gln Lys Asn Ile Glu Thr Ser Gly Ala Arg Cys Glu Gln Pro20 25 30Lys Arg Leu Arg Lys Lys Lys Phe Ala Thr Leu Pro Pro Asn Lys Phe35 40 45Lys Cys Lys Gly Ser Glu Ser Phe Val Ser Met Tyr Ala Asp Ser Cys50 55 60Phe Ile Asp Ser Ile Cys Pro Thr Gln Cys Asp Cys Tyr Gly Thr Thr65 70 75 80Val Asp Cys Asn Lys Arg Gly Leu Asn Thr Ile Pro Thr Ser Ile Pro85 90 95Arg Phe Ala Thr Gln Leu Leu Leu Ser Gly Asn Asn Ile Ser Thr Val100 105 110Asp Leu Asn Ser Asn Ile His Val Leu Glu Asn Leu Glu Xaa Leu Asp115 120 125Leu Ser Asn Asn His Ile Thr Phe Ile Asn Asp Lys Ser Phe Glu Lys130 135 140Leu Ser Lys Leu Arg Glu Leu Xaa Leu Asn Asp145 150 1559735PRTCaenorhabditis elegans 9Ser Asn Lys Asn Leu Thr Ser Phe Pro Ser Arg Ile Pro Phe Asp Thr1 5 10 15Thr Glu Leu Tyr Leu Asp Ala Asn Tyr Ile Asn Glu Ile Pro Ala His20 25 30Asp Leu Asn Arg Leu Tyr Ser Leu Thr Lys Leu Asp Leu Ser His Asn35 40 45Arg Leu Ile Ser Leu Glu Asn Asn Thr Phe Ser Asn Leu Thr Arg Leu50 55 60Ser Thr Leu Ile Ile Ser Tyr Asn Lys Leu Arg Cys Leu Gln Pro Leu65 70 75 80Ala Phe Asn Gly Leu Asn Ala Leu Arg Ile Leu Ser Leu His Gly Asn85 90 95Asp Ile Ser Phe Leu Pro Gln Ser Ala Phe Ser Asn Leu Thr Ser Ile100 105 110Thr His Ile Ala Val Gly Ser Asn Ser Leu Tyr Cys Asp Cys Asn Met115 120 125Ala Trp Phe Ser Lys Trp Ile Lys Ser Lys Phe Ile Glu Ala Gly Ile130 135 140Ala Arg Cys Glu Tyr Pro Asn Thr Val Ser Asn Gln Leu Leu Leu Thr145 150 155 160Ala Gln Pro Tyr Gln Phe Thr Cys Asp Ser Lys Val Pro Thr Lys Leu165 170 175Ala Thr Lys Cys Asp Leu Cys Leu Asn Ser Pro Cys Lys Asn Asn Ala180 185 190Ile Cys Glu Thr Thr Ser Ser Arg Lys Tyr Thr Cys Asn Cys Thr Pro195 200 205Gly Phe Tyr Gly Val His Cys Glu Asn Gln Ile Asp Ala Cys Tyr Gly210 215 220Ser Pro Cys Leu Asn Asn Ala Thr Cys Lys Val Ala Gln Ala Gly Arg225 230 235 240Phe Asn Cys Tyr Cys Asn Lys Gly Phe Glu Gly Asp Tyr Cys Glu Lys245 250 255Asn Ile Asp Asp Cys Val Asn Ser Lys Cys Glu Asn Gly Gly Lys Cys260 265 270Val Asp Leu Val Arg Phe Cys Ser Glu Glu Leu Lys Asn Phe Gln Ser275 280 285Phe Gln Ile Asn Ser Tyr Arg Cys Asp Cys Pro Met Glu Tyr Glu Gly290 295 300Lys His Cys Glu Asp Lys Leu Glu Tyr Cys Thr Lys Lys Leu Asn Pro305 310 315 320Cys Glu Asn Asn Gly Lys Cys Ile Pro Ile Asn Gly Ser Tyr Ser Cys325 330 335Met Cys Ser Pro Gly Phe Thr Gly Asn Asn Cys Glu Thr Asn Ile Asp340 345 350Asp Cys Lys Asn Val Glu Cys Gln Asn Gly Gly Ser Cys Val Asp Gly355 360 365Ile Leu Ser Tyr Asp Cys Leu Cys Arg Pro Gly Tyr Ala Gly Gln Tyr370 375 380Cys Glu Ile Pro Pro Met Met Asp Met Glu Tyr Gln Lys Thr Asp Ala385 390 395 400Cys Gln Gln Ser Ala Cys Gly Gln Gly Glu Cys Val Ala Ser Gln Asn405 410 415Ser Ser Asp Phe Thr Cys Lys Cys His Glu Gly Phe Ser Gly Pro Ser420 425 430Cys Asp Arg Gln Met Ser Val Gly Phe Lys Asn Pro Gly Ala Tyr Leu435 440 445Ala Leu Asp Pro Leu Ala Ser Asp Gly Thr Ile Thr Met Thr Leu Arg450 455 460Thr Thr Ser Lys Ile Gly Ile Leu Leu Tyr Tyr Gly Asp Asp His Phe465 470 475 480Val Ser Ala Glu Leu Tyr Asp Gly Arg Val Lys Leu Val Tyr Tyr Ile485 490 495Gly Asn Phe Pro Ala Ser His Met Tyr Ser Ser Val Lys Val Asn Asp500 505 510Gly Leu Pro His Arg Ile Ser Ile Arg Thr Ser Glu Arg Lys Cys Phe515 520 525Leu Gln Ile Asp Lys Asn Pro Val Gln Ile Val Glu Asn Ser Gly Lys530 535 540Ser Asp Gln Leu Ile Thr Lys Gly Lys Glu Met Leu Tyr Ile Gly Gly545 550 555 560Leu Pro Ile Glu Lys Ser Gln Asp Ala Lys Arg Arg Phe His Val Lys565 570 575Asn Ser Glu Ser Leu Lys Gly Cys Ile Ser Ser Ile Thr Ile Asn Glu580 585 590Val Pro Ile Asn Leu Gln Gln Ala Leu Glu Asn Val Asn Thr Glu Gln595 600 605Ser Cys Ser Ala Thr Val Asn Phe Cys Ala Gly Ile Asp Cys Gly Asn610 615 620Gly Lys Cys Thr Asn Asn Ala Leu Ser Pro Lys Gly Tyr Met Cys Gln625 630 635 640Cys Asp Ser His Phe Ser Gly Glu His Cys Asp Glu Lys Arg Ile Lys645 650 655Cys Asp Lys Gln Lys Phe Arg Arg His His Ile Glu Asn Glu Cys Arg660 665 670Ser Val Asp Arg Ile Lys Ile Ala Glu Cys Asn Gly Tyr Cys Gly Gly675 680 685Glu Gln Asn Cys Cys Thr Ala Val Lys Lys Lys Gln Arg Lys Val Lys690 695 700Met Ile Cys Lys Asn Gly Thr Thr Lys Ile Ser Thr Val His Ile Ile705 710 715 720Arg Gln Cys Gln Cys Glu Pro Thr Lys Ser Val Leu Ser Glu Lys725 730 73510154PRTmouse 10Asp Pro Leu Pro Val His His Arg Cys Glu Cys Met Leu Gly Tyr Thr1 5 10 15Gly Asp Asn Cys Ser Glu Asn Gln Asp Asp Cys Lys Asp His Lys Cys20 25 30Gln Asn Gly Ala Gln Cys Val Asp Glu Val Asn Ser Tyr Ala Cys Leu35 40 45Cys Val Glu Gly Tyr Ser Gly Gln Leu Cys Glu Ile Pro Pro Ala Pro50 55 60Arg Ser Ser Cys Glu Gly Thr Glu Cys Gln Asn Gly Ala Asn Cys Val65 70 75 80Asp Gln Gly Ser Arg Pro Val Cys Gln Cys Leu Pro Gly Phe Gly Gly85 90 95Pro Glu Cys Glu Lys Leu Leu Ser Val Asn Phe Val Asp Arg Asp Thr100 105 110Tyr Leu Gln Phe Thr Asp Leu Gln Asn Trp Pro Arg Ala Asn Ile Thr115 120 125Leu Gln Val Ser Thr Ala Glu Asp Asn Gly Ile Leu Leu Tyr Asn Gly130 135 140Asp Asn Asp His Ile Ala Val Glu Leu Tyr145 15011110PRTmouse 11Ala Phe Lys Cys His His Gly Gln Cys His Ile Ser Asp Arg Gly Glu1 5 10 15Pro Tyr Cys Leu Cys Gln Pro Gly Phe Ser Gly His His Cys Glu Gln20 25 30Glu Asn Pro Cys Met Gly Glu Ile Val Arg Glu Ala Ile Arg Arg Gln35 40 45Lys Asp Tyr Ala Ser Cys Ala Thr Ala Ser Lys Val Pro Ile Met Glu50 55 60Cys Arg Gly Gly Cys Gly Thr Thr Cys Cys Gln Pro Ile Arg Ser Lys65 70 75 80Arg Arg Lys Tyr Val Phe Gln Cys Thr Asp Gly Ser Ser Phe Val Glu85 90 95Glu Val Glu Arg His Leu Glu Cys Gly Cys Arg Ala Cys Ser100 105 11012134PRTmouse 12His Leu Arg Val Leu Gln Leu Met Glu Asn Arg Ile Ser Thr Ile Glu1 5 10 15Arg Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn20 25 30Arg Asn Asn Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala35 40 45Arg Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro50 55 60Arg Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp65 70 75 80Tyr Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg85 90 95Asp Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser100 105 110Val Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His115 120 125Ser Asn Asn Leu Tyr Cys13013104PRTmouse 13Asn Asn Asp Asp Cys Val Gly His Lys Cys Arg His Gly Ala Gln Cys1 5 10 15Val Asp Glu Val Asn Gly Tyr Thr Cys Ile Cys Pro Gln Gly Phe Ser20 25 30Gly Leu Phe Cys Glu His Pro Pro Pro Met Val Leu Leu Gln Thr Ser35 40 45Pro Cys Asp Gln Tyr Glu Cys Gln Asn Gly Ala Gln Cys Ile Val Val50 55 60Gln Gln Glu Pro Thr Cys Arg Cys Pro Pro Gly Phe Ala Gly Pro Arg65 70 75 80Cys Glu Lys Leu Ile Thr Val Asn Phe Val Gly Lys Asp Ser Tyr Val85 90 95Glu Leu Ala Ser Ala Lys Val Arg10014243PRTmouse 14Ile Leu Asp Val Ala Ser Leu Arg Gln Ala Pro Gly Glu Asn Gly Thr1 5 10 15Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu Leu Gln20 25 30Asp Phe Arg Lys Met Pro Met Gln Thr Gly Ile Leu Pro Gly Cys Glu35 40 45Pro Cys His Lys Lys Val Cys Ala His Gly Cys Cys Gln Pro Ser Ser50 55 60Gln Ser Gly Phe Thr Cys Glu Cys Glu Glu Gly Trp Met Gly Pro Leu65 70 75 80Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys Val His85 90 95Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys Cys Leu100 105 110Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu Phe Asn115 120 125Pro Cys Gln Met Ile Lys Cys Lys His Gly Lys Cys Arg Leu Ser Gly130 135 140Val Gly Gln Pro Tyr Cys Glu Cys Asn Ser Gly Phe Thr Gly Asp Ser145 150 155 160Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp Tyr Tyr165 170 175Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys Val Ser180 185 190Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys Gly Pro195 200 205Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp Gly Ser210 215 220Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys Ala Arg225 230 235 240Cys Ala Ser151395PRTDrosophila melanogaster 15Met His Pro Met His Pro Glu Asn His Ala Ile Ala Arg Ser Thr Ser1 5 10 15Thr Thr Asn Asn Pro Ser Arg Ser Arg Ser Ser Arg Met Trp Leu Leu20 25 30Pro Ala Trp Leu Leu Leu Val Leu Val Ala Ser Asn Gly Leu Pro Ala35 40 45Val Arg Gly Gln Tyr Gln Ser Pro Arg Ile Ile Glu His Pro Thr Asp50 55 60Leu Val Val Lys Lys Asn Glu Pro Ala Thr Leu Asn Cys Lys Val Glu65 70 75 80Gly Lys Pro Glu Pro Thr Ile Glu Trp Phe Lys Asp Gly Glu Pro Val85 90 95Ser Thr Asn Glu Lys Lys Ser His Arg Val Gln Phe Lys Asp Gly Ala100 105 110Leu Phe Phe Tyr Arg Thr Met Gln Gly Lys Lys Glu Gln Asp Gly Gly115 120 125Glu Tyr Trp Cys Val Ala Lys Asn Arg Val Gly Gln Ala Val Ser Arg130 135 140His Ala Ser Leu Gln Ile Ala Val Leu Arg Asp Asp Phe Arg Val Glu145 150 155 160Pro Lys Asp Thr Arg Val Ala Lys Gly Glu Thr Ala Leu Leu Glu Cys165 170 175Gly Pro Pro Lys Gly Ile Pro Glu Pro Thr Leu Ile Trp Ile Lys Asp180

185 190Gly Val Pro Leu Asp Asp Leu Lys Ala Met Ser Phe Gly Ala Ser Ser195 200 205Arg Val Arg Ile Val Asp Gly Gly Asn Leu Leu Ile Ser Asn Val Glu210 215 220Pro Ile Asp Glu Gly Asn Tyr Lys Cys Ile Ala Gln Asn Leu Val Gly225 230 235 240Thr Arg Glu Ser Ser Tyr Ala Lys Leu Ile Val Gln Val Lys Pro Tyr245 250 255Phe Met Lys Glu Pro Lys Asp Gln Val Met Leu Tyr Gly Gln Thr Ala260 265 270Thr Phe His Cys Ser Val Gly Gly Asp Pro Pro Pro Lys Val Leu Trp275 280 285Lys Lys Glu Glu Gly Asn Ile Pro Val Ser Arg Ala Arg Ile Leu His290 295 300Asp Glu Lys Ser Leu Glu Ile Ser Asn Ile Thr Pro Thr Asp Glu Gly305 310 315 320Thr Tyr Val Cys Glu Ala His Asn Asn Val Gly Gln Ile Ser Ala Arg325 330 335Ala Ser Leu Ile Val His Ala Pro Pro Asn Phe Thr Lys Arg Pro Ser340 345 350Asn Lys Lys Val Gly Leu Asn Gly Val Val Gln Leu Pro Cys Met Ala355 360 365Ser Gly Asn Pro Pro Pro Ser Val Phe Trp Thr Lys Glu Gly Val Ser370 375 380Thr Leu Met Phe Pro Asn Ser Ser His Gly Arg Gln Tyr Val Ala Ala385 390 395 400Asp Gly Thr Leu Gln Ile Thr Asp Val Arg Gln Glu Asp Glu Gly Tyr405 410 415Tyr Val Cys Ser Ala Phe Ser Val Val Asp Ser Ser Thr Val Arg Val420 425 430Phe Leu Gln Val Ser Ser Val Asp Glu Arg Pro Pro Pro Ile Ile Gln435 440 445Ile Gly Pro Ala Asn Gln Thr Leu Pro Lys Gly Ser Val Ala Thr Leu450 455 460Pro Cys Arg Ala Thr Gly Asn Pro Ser Pro Arg Ile Lys Trp Phe His465 470 475 480Asp Gly His Ala Val Gln Ala Gly Asn Arg Tyr Ser Ile Ile Gln Gly485 490 495Ser Ser Leu Arg Val Asp Asp Leu Gln Leu Ser Asp Ser Gly Thr Tyr500 505 510Thr Cys Thr Ala Ser Gly Glu Arg Gly Glu Thr Ser Trp Ala Ala Thr515 520 525Leu Thr Val Glu Lys Pro Gly Ser Thr Ser Leu His Arg Ala Ala Asp530 535 540Pro Ser Thr Tyr Pro Ala Pro Pro Gly Thr Pro Lys Val Leu Asn Val545 550 555 560Ser Arg Thr Ser Ile Ser Leu Arg Trp Ala Lys Ser Gln Glu Lys Pro565 570 575Gly Ala Val Gly Pro Ile Ile Gly Tyr Thr Val Glu Tyr Phe Ser Pro580 585 590Asp Leu Gln Thr Gly Trp Ile Val Ala Ala His Arg Val Gly Asp Thr595 600 605Gln Val Thr Ile Ser Gly Leu Thr Pro Gly Thr Ser Tyr Val Phe Leu610 615 620Val Arg Ala Glu Asn Thr Gln Gly Ile Ser Val Pro Ser Gly Leu Ser625 630 635 640Asn Val Ile Lys Thr Ile Glu Ala Asp Phe Asp Ala Ala Ser Ala Asn645 650 655Asp Leu Ser Ala Ala Arg Thr Leu Leu Thr Gly Lys Ser Val Glu Leu660 665 670Ile Asp Ala Ser Ala Ile Asn Ala Ser Ala Val Arg Leu Glu Trp Met675 680 685Leu His Val Ser Ala Asp Glu Lys Tyr Val Glu Gly Leu Arg Ile His690 695 700Tyr Lys Asp Ala Ser Val Pro Ser Ala Gln Tyr His Ser Ile Thr Val705 710 715 720Met Asp Ala Ser Ala Glu Ser Phe Val Val Gly Asn Leu Lys Lys Tyr725 730 735Thr Lys Tyr Glu Phe Phe Leu Thr Pro Phe Phe Glu Thr Ile Glu Gly740 745 750Gln Pro Ser Asn Ser Lys Thr Ala Leu Thr Tyr Glu Asp Val Pro Ser755 760 765Ala Pro Pro Asp Asn Ile Gln Ile Gly Met Tyr Asn Gln Thr Ala Gly770 775 780Trp Val Arg Trp Thr Pro Pro Pro Ser Gln His His Asn Gly Asn Leu785 790 795 800Tyr Gly Tyr Lys Ile Glu Val Ser Ala Gly Asn Thr Met Lys Val Leu805 810 815Ala Asn Met Thr Leu Asn Ala Thr Thr Thr Ser Val Leu Leu Asn Asn820 825 830Leu Thr Thr Gly Ala Val Tyr Ser Val Arg Leu Asn Ser Phe Thr Lys835 840 845Ala Gly Asp Gly Pro Tyr Ser Lys Pro Ile Ser Leu Phe Met Asp Pro850 855 860Thr His His Val His Pro Pro Arg Ala His Pro Ser Gly Thr His Asp865 870 875 880Gly Arg His Glu Gly Gln Asp Leu Thr Tyr His Asn Asn Gly Asn Ile885 890 895Pro Pro Gly Asp Ile Asn Pro Thr Thr His Lys Lys Thr Thr Asp Tyr900 905 910Leu Ser Gly Pro Trp Leu Met Val Leu Val Cys Ile Val Leu Leu Val915 920 925Leu Val Ile Ser Ala Ala Ile Ser Met Val Tyr Phe Lys Arg Lys His930 935 940Gln Met Thr Lys Glu Leu Gly His Leu Ser Val Val Ser Asp Asn Glu945 950 955 960Ile Thr Ala Leu Asn Ile Asn Ser Lys Glu Ser Leu Trp Ile Asp His965 970 975His Arg Gly Trp Arg Thr Ala Asp Thr Asp Lys Asp Ser Gly Leu Ser980 985 990Glu Ser Lys Leu Leu Ser His Val Asn Ser Ser Gln Ser Asn Tyr Asn995 1000 1005Asn Ser Asp Gly Gly Thr Asp Tyr Ala Glu Val Asp Thr Arg Asn Leu1010 1015 1020Thr Thr Phe Tyr Asn Cys Arg Lys Ser Pro Asp Asn Pro Thr Pro Tyr1025 1030 1035 1040Ala Thr Thr Met Ile Ile Gly Thr Ser Ser Ser Glu Thr Cys Thr Lys1045 1050 1055Thr Thr Ser Ile Ser Ala Asp Lys Asp Ser Gly Thr His Ser Pro Tyr1060 1065 1070Ser Asp Ala Phe Ala Gly Gln Val Pro Ala Val Pro Val Val Lys Ser1075 1080 1085Asn Tyr Leu Gln Tyr Pro Val Glu Pro Ile Asn Trp Ser Glu Phe Leu1090 1095 1100Pro Pro Pro Pro Glu His Pro Pro Pro Ser Ser Thr Tyr Gly Tyr Ala1105 1110 1115 1120Gln Gly Ser Pro Glu Ser Ser Arg Lys Ser Ser Lys Ser Ala Gly Ser1125 1130 1135Gly Ile Ser Thr Asn Gln Ser Ile Leu Asn Ala Ser Ile His Ser Ser1140 1145 1150Ser Ser Gly Gly Phe Ser Ala Trp Gly Val Ser Pro Gln Tyr Ala Val1155 1160 1165Ala Cys Pro Pro Glu Asn Val Tyr Ser Asn Pro Leu Ser Ala Val Ala1170 1175 1180Gly Gly Thr Gln Asn Arg Tyr Gln Ile Thr Pro Thr Asn Gln His Pro1185 1190 1195 1200Pro Gln Leu Pro Ala Tyr Phe Ala Thr Thr Gly Pro Gly Gly Ala Val1205 1210 1215Pro Pro Asn His Leu Pro Phe Ala Thr Gln Arg His Ala Ala Ser Glu1220 1225 1230Tyr Gln Ala Gly Leu Asn Ala Ala Arg Cys Ala Gln Ser Arg Ala Cys1235 1240 1245Asn Ser Cys Asp Ala Leu Ala Thr Pro Ser Pro Met Gln Pro Pro Pro1250 1255 1260Pro Val Pro Val Pro Glu Gly Trp Tyr Gln Pro Val His Pro Asn Ser1265 1270 1275 1280His Pro Met His Pro Thr Ser Ser Asn His Gln Ile Tyr Gln Cys Ser1285 1290 1295Ser Glu Cys Ser Asp His Ser Arg Ser Ser Gln Ser His Lys Arg Gln1300 1305 1310Leu Gln Leu Glu Glu His Gly Ser Ser Ala Lys Gln Arg Gly Gly His1315 1320 1325His Arg Arg Arg Ala Pro Val Val Gln Pro Cys Met Glu Ser Glu Asn1330 1335 1340Glu Asn Met Leu Ala Glu Tyr Glu Gln Arg Gln Tyr Thr Ser Asp Cys1345 1350 1355 1360Cys Asn Ser Ser Arg Glu Gly Asp Thr Cys Ser Cys Ser Glu Gly Ser1365 1370 1375Cys Leu Tyr Ala Glu Ala Gly Glu Pro Ala Pro Arg Gln Met Thr Ala1380 1385 1390Lys Asn Thr1395161381PRTDrosophila melanogaster 16Gly Glu Asn Pro Arg Ile Ile Glu His Pro Met Asp Thr Thr Val Pro1 5 10 15Lys Asn Asp Pro Phe Thr Phe Asn Cys Gln Ala Glu Gly Asn Pro Thr20 25 30Pro Thr Ile Gln Trp Phe Lys Asp Gly Arg Glu Leu Lys Thr Asp Thr35 40 45Gly Ser His Arg Ile Met Leu Pro Ala Gly Gly Leu Phe Phe Leu Lys50 55 60Val Ile His Ser Arg Arg Glu Ser Asp Ala Gly Thr Tyr Trp Cys Glu65 70 75 80Ala Lys Asn Glu Phe Gly Val Ala Arg Ser Arg Asn Ala Thr Leu Gln85 90 95Val Ala Val Leu Arg Asp Glu Phe Arg Leu Glu Pro Ala Asn Thr Arg100 105 110Val Ala Gln Gly Glu Val Ala Leu Met Glu Cys Gly Ala Pro Arg Gly115 120 125Ser Pro Glu Pro Gln Ile Ser Trp Arg Lys Asn Gly Gln Thr Leu Asn130 135 140Leu Val Gly Asn Lys Arg Ile Arg Ile Val Asp Gly Gly Asn Leu Ala145 150 155 160Ile Gln Glu Ala Arg Gln Ser Asp Asp Gly Arg Tyr Gln Cys Val Val165 170 175Lys Asn Val Val Gly Thr Arg Glu Ser Ala Thr Ala Phe Leu Lys Val180 185 190His Val Arg Pro Phe Leu Ile Arg Gly Pro Gln Asn Gln Thr Ala Val195 200 205Val Gly Ser Ser Val Val Phe Gln Cys Arg Ile Gly Gly Asp Pro Leu210 215 220Pro Asp Val Leu Trp Arg Arg Thr Ala Ser Gly Gly Asn Met Pro Leu225 230 235 240Arg Lys Phe Ser Trp Leu His Ser Ala Ser Gly Arg Val His Val Leu245 250 255Glu Asp Arg Ser Leu Lys Leu Asp Asp Val Thr Leu Glu Asp Met Gly260 265 270Glu Tyr Thr Cys Glu Ala Asp Asn Ala Val Gly Gly Ile Thr Ala Thr275 280 285Gly Ile Leu Thr Val His Ala Pro Pro Lys Phe Val Ile Arg Pro Lys290 295 300Asn Gln Leu Val Glu Ile Gly Asp Glu Val Leu Phe Glu Cys Gln Ala305 310 315 320Asn Gly His Pro Arg Pro Thr Leu Tyr Trp Ser Val Glu Gly Asn Ser325 330 335Ser Leu Leu Leu Pro Gly Tyr Arg Asp Gly Arg Met Glu Val Thr Leu340 345 350Thr Pro Glu Gly Arg Ser Val Leu Ser Ile Ala Arg Phe Ala Arg Glu355 360 365Asp Ser Gly Lys Val Val Thr Cys Asn Ala Leu Asn Ala Val Gly Ser370 375 380Val Ser Ser Arg Thr Val Val Ser Val Asp Thr Gln Phe Glu Leu Pro385 390 395 400Pro Pro Ile Ile Glu Gln Gly Pro Val Asn Gln Thr Leu Pro Val Lys405 410 415Ser Ile Val Val Leu Pro Cys Arg Thr Leu Gly Thr Pro Val Pro Gln420 425 430Val Ser Trp Tyr Leu Asp Gly Ile Pro Ile Asp Val Gln Glu His Glu435 440 445Arg Arg Asn Leu Ser Asp Ala Gly Ala Leu Thr Ile Ser Asp Leu Gln450 455 460Arg His Glu Asp Glu Gly Leu Tyr Thr Cys Val Ala Ser Asn Arg Asn465 470 475 480Gly Lys Ser Ser Trp Ser Gly Tyr Leu Arg Leu Asp Thr Pro Thr Asn485 490 495Pro Asn Ile Lys Phe Phe Arg Ala Pro Glu Leu Ser Thr Tyr Pro Gly500 505 510Pro Pro Gly Lys Pro Gln Met Val Glu Lys Gly Glu Asn Ser Val Thr515 520 525Leu Ser Trp Thr Arg Ser Asn Lys Val Gly Gly Ser Ser Leu Val Gly530 535 540Tyr Val Ile Glu Met Phe Gly Lys Asn Glu Thr Asp Gly Trp Val Ala545 550 555 560Val Gly Thr Arg Val Gln Asn Thr Thr Phe Thr Gln Thr Gly Leu Leu565 570 575Pro Gly Val Asn Tyr Phe Phe Leu Ile Arg Ala Glu Asn Ser His Gly580 585 590Leu Ser Leu Pro Ser Pro Met Ser Glu Pro Ile Thr Val Gly Thr Arg595 600 605Tyr Phe Asn Ser Gly Leu Asp Leu Ser Glu Ala Arg Ala Ser Leu Leu610 615 620Ser Gly Asp Val Val Glu Leu Ser Asn Ala Ser Val Val Asp Ser Thr625 630 635 640Ser Met Lys Leu Thr Trp Gln Ile Ile Asn Gly Lys Tyr Val Glu Gly645 650 655Phe Tyr Val Tyr Ala Arg Gln Leu Pro Asn Pro Ile Val Asn Asn Pro660 665 670Ala Pro Val Thr Ser Asn Thr Asn Pro Leu Leu Gly Ser Thr Ser Thr675 680 685Ser Ala Ser Ala Ser Ala Ser Ala Ser Ala Leu Ile Ser Thr Lys Pro690 695 700Asn Ile Ala Ala Ala Gly Lys Arg Asp Gly Glu Thr Asn Gln Ser Gly705 710 715 720Gly Gly Ala Pro Thr Pro Leu Asn Thr Lys Tyr Arg Met Leu Thr Ile725 730 735Leu Asn Gly Gly Gly Ala Ser Ser Cys Thr Ile Thr Gly Leu Val Gln740 745 750Tyr Thr Leu Tyr Glu Phe Phe Ile Val Pro Phe Tyr Lys Ser Val Glu755 760 765Gly Lys Pro Ser Asn Ser Arg Ile Ala Arg Thr Leu Glu Asp Val Pro770 775 780Ser Glu Ala Pro Tyr Gly Met Glu Ala Leu Leu Leu Asn Ser Ser Ala785 790 795 800Val Phe Leu Lys Trp Lys Ala Pro Glu Leu Lys Asp Arg His Gly Val805 810 815Leu Leu Asn Tyr His Val Ile Val Arg Gly Ile Asp Thr Ala His Asn820 825 830Phe Ser Arg Ile Leu Thr Asn Val Thr Ile Asp Ala Ala Ser Pro Thr835 840 845Leu Val Leu Ala Asn Leu Thr Glu Gly Val Met Tyr Thr Val Gly Val850 855 860Ala Ala Gly Asn Asn Ala Gly Val Gly Pro Tyr Cys Val Pro Ala Thr865 870 875 880Leu Arg Leu Asp Pro Ile Thr Lys Arg Leu Asp Pro Phe Ile Asn Gln885 890 895Arg Asp His Val Asn Asp Val Leu Thr Gln Pro Trp Phe Ile Ile Leu900 905 910Leu Gly Ala Ile Leu Ala Val Leu Met Leu Ser Phe Gly Ala Met Val915 920 925Phe Val Lys Arg Lys His Met Met Met Lys Gln Ser Ala Leu Asn Thr930 935 940Met Arg Gly Asn His Thr Ser Asp Val Leu Lys Met Pro Ser Leu Ser945 950 955 960Ala Arg Asn Gly Asn Gly Tyr Trp Leu Asp Ser Ser Thr Gly Gly Met965 970 975Val Trp Arg Pro Ser Pro Gly Gly Asp Ser Leu Glu Met Gln Lys Asp980 985 990His Ile Ala Asp Tyr Ala Pro Val Cys Gly Ala Pro Gly Ser Pro Ala995 1000 1005Gly Gly Gly Thr Ser Ser Gly Gly Ser Gly Gly Ala Gly Ser Gly Ala1010 1015 1020Ser Gly Gly Asp Asp Ile His Gly Gly His Gly Ser Glu Arg Asn Gln1025 1030 1035 1040Gln Arg Tyr Val Gly Glu Tyr Ser Asn Ile Pro Thr Asp Tyr Ala Glu1045 1050 1055Val Ser Ser Phe Gly Lys Ala Pro Ser Glu Tyr Gly Arg His Gly Asn1060 1065 1070Ala Ser Pro Ala Pro Tyr Ala Thr Ser Ser Ile Leu Ser Pro His Gln1075 1080 1085Gln Gln Gln Gln Gln Gln Pro Arg Tyr Gln Gln Arg Pro Val Pro Gly1090 1095 1100Tyr Gly Leu Gln Arg Pro Met His Pro His Tyr Gln Gln Gln Gln His1105 1110 1115 1120Gln Gln Gln Gln Ala Gln Gln Thr His Gln Gln His Gln Ala Leu Gln1125 1130 1135Gln His Gln Gln Leu Pro Pro Ser Asn Ile Tyr Gln Gln Met Ser Thr1140 1145 1150Thr Ser Glu Ile Tyr Pro Thr Asn Thr Gly Pro Ser Arg Ser Val Tyr1155 1160 1165Ser Glu Gln Tyr Tyr Tyr Pro Lys Asp Lys Gln Arg His Ile His Ile1170 1175 1180Thr Glu Asn Lys Leu Ser Asn Cys His Thr Tyr Glu Ala Ala Pro Gly1185 1190 1195 1200Ala Lys Gln Ser Ser Pro Ile Ser Ser Gln Phe Ala Ser Val Arg Arg1205 1210 1215Gln Gln Leu Pro Pro Asn Cys Ser Ile Gly Arg Glu Ser Ala Arg Phe1220 1225 1230Lys Val Leu Asn Thr Asp Gln Gly Lys Asn Gln Gln Asn Leu Leu Asp1235 1240 1245Leu Asp Gly Ser Ser Met Cys Tyr Asn Gly Leu Ala Asp Ser Gly Cys1250 1255 1260Gly Gly Ser Pro Ser Pro Met Ala Met Leu Met Ser His Glu Asp Glu1265 1270 1275 1280His Ala Leu Tyr His Thr Ala Asp Gly Asp Leu Asp Asp Met Glu Arg1285 1290 1295Leu Tyr Val Lys Val Asp Glu Gln Gln Pro Pro Gln Gln Gln Gln Gln1300 1305 1310Leu Ile Pro Leu Val Pro Gln His Pro Ala Glu Gly His Leu Gln Ser1315 1320 1325Trp Arg Asn Gln Ser Thr Arg Ser Ser Arg Lys Asn Gly Gln Glu Cys1330 1335 1340Ile Lys Glu Pro Ser Glu Leu Ile Tyr Ala Pro Gly Ser Val Ala Ser1345 1350 1355 1360Glu Arg Ser Leu Leu Ser Asn Ser Gly Ser Gly Thr Ser Ser Gln Pro1365 1370 1375Ala Gly His Asn Val1380171297PRTCaenorhabditis elegans 17Met Tyr Tyr Leu Gly Phe Tyr His Thr His Thr His Thr His Thr Tyr1 5 10 15Ile Asn Phe Asp Lys Ile Pro Asn Ala Ser Asn Leu Ala Pro Val Ile20 25 30Ile Glu His Pro Ile Asp Val Val Val Ser Arg Gly Ser Pro Ala Thr35 40 45Leu Asn Cys Gly Ala

Lys Pro Ser Thr Ala Lys Ile Thr Trp Tyr Lys50 55 60Asp Gly Gln Pro Val Ile Thr Asn Lys Glu Gln Val Asn Ser His Arg65 70 75 80Ile Val Leu Asp Thr Gly Ser Leu Phe Leu Leu Lys Val Asn Ser Gly85 90 95Lys Asn Gly Lys Asp Ser Asp Ala Gly Ala Tyr Tyr Cys Val Ala Ser100 105 110Asn Glu His Gly Glu Val Lys Ser Asn Glu Gly Ser Leu Lys Leu Ala115 120 125Met Leu Arg Glu Asp Phe Arg Val Arg Pro Arg Thr Val Gln Ala Leu130 135 140Gly Gly Glu Met Ala Val Leu Glu Cys Ser Pro Pro Arg Gly Phe Pro145 150 155 160Glu Pro Val Val Ser Trp Arg Lys Asp Asp Lys Glu Leu Arg Ile Gln165 170 175Asp Met Pro Arg Tyr Thr Leu His Ser Asp Gly Asn Leu Ile Ile Asp180 185 190Pro Val Asp Arg Ser Asp Ser Gly Thr Tyr Gln Cys Val Ala Asn Asn195 200 205Met Val Gly Glu Arg Val Ser Asn Pro Ala Arg Leu Ser Val Phe Glu210 215 220Lys Pro Lys Phe Glu Gln Glu Pro Lys Asp Met Thr Val Asp Val Gly225 230 235 240Ala Ala Val Leu Phe Asp Cys Arg Val Thr Gly Asp Pro Gln Pro Gln245 250 255Ile Thr Trp Lys Arg Lys Asn Glu Pro Met Pro Val Thr Arg Ala Tyr260 265 270Ile Ala Lys Asp Asn Arg Gly Leu Arg Ile Glu Arg Val Gln Pro Ser275 280 285Asp Glu Gly Glu Tyr Val Cys Tyr Ala Arg Asn Pro Ala Gly Thr Leu290 295 300Glu Ala Ser Ala His Leu Arg Val Gln Ala Pro Pro Ser Phe Gln Thr305 310 315 320Lys Pro Ala Asp Gln Ser Val Pro Ala Gly Gly Thr Ala Thr Phe Glu325 330 335Cys Thr Leu Val Gly Gln Pro Ser Pro Ala Tyr Phe Trp Ser Lys Glu340 345 350Gly Gln Gln Asp Leu Leu Phe Pro Ser Tyr Val Ser Ala Asp Gly Arg355 360 365Thr Lys Val Ser Pro Thr Gly Thr Leu Thr Ile Glu Glu Val Arg Gln370 375 380Val Asp Glu Gly Ala Tyr Val Cys Ala Gly Met Asn Ser Ala Gly Ser385 390 395 400Ser Leu Ser Lys Ala Ala Leu Lys Ala Thr Phe Glu Thr Lys Gly Arg405 410 415Val Gln Lys Lys Lys Ser Lys Met Gly Lys Gln Lys Gln Lys Asn Val420 425 430Gln Ser Ile Ile Lys Tyr Leu Ile Ser Ala Val Thr Gly Asn Thr Pro435 440 445Ala Lys Pro Pro Pro Thr Ile Glu His Gly His Gln Asn Gln Thr Leu450 455 460Met Val Gly Ser Ser Ala Ile Leu Pro Cys Gln Ala Ser Gly Lys Pro465 470 475 480Thr Pro Gly Ile Ser Trp Leu Arg Asp Gly Leu Pro Ile Asp Ile Thr485 490 495Asp Ser Arg Ile Ser Gln His Ser Thr Gly Ser Leu His Ile Ala Asp500 505 510Leu Lys Lys Pro Asp Thr Gly Val Tyr Thr Cys Ile Ala Lys Asn Glu515 520 525Asp Gly Glu Ser Thr Trp Ser Ala Ser Leu Thr Val Glu Asp His Thr530 535 540Ser Asn Ala Gln Phe Val Arg Met Pro Asp Pro Ser Asn Phe Pro Ser545 550 555 560Ser Pro Thr Gln Pro Ile Ile Val Asn Val Thr Asp Thr Glu Val Glu565 570 575Leu His Trp Asn Ala Pro Ser Thr Ser Gly Ala Gly Pro Ile Thr Gly580 585 590Tyr Ile Ile Gln Tyr Tyr Ser Pro Asp Leu Gly Gln Thr Trp Phe Asn595 600 605Ile Pro Asp Tyr Val Ala Ser Thr Glu Tyr Arg Ile Lys Gly Leu Lys610 615 620Pro Ser His Ser Tyr Met Phe Val Ile Arg Ala Glu Asn Glu Lys Gly625 630 635 640Ile Gly Thr Pro Ser Val Ser Ser Ala Leu Val Thr Thr Ser Lys Pro645 650 655Ala Ala Gln Val Ala Leu Ser Asp Lys Asn Lys Met Asp Met Ala Ile660 665 670Ala Glu Lys Arg Leu Thr Ser Glu Gln Leu Ile Lys Leu Glu Glu Val675 680 685Lys Thr Ile Asn Ser Thr Ala Val Arg Leu Phe Trp Lys Lys Arg Lys690 695 700Leu Glu Glu Leu Ile Asp Gly Tyr Tyr Ile Lys Trp Arg Gly Pro Pro705 710 715 720Arg Thr Asn Asp Asn Gln Tyr Val Asn Val Thr Ser Pro Ser Thr Glu725 730 735Asn Tyr Val Val Ser Asn Leu Met Pro Phe Thr Asn Tyr Glu Phe Phe740 745 750Val Ile Pro Tyr His Ser Gly Val His Ser Ile His Gly Ala Pro Ser755 760 765Asn Ser Met Asp Val Leu Thr Ala Glu Ala Pro Pro Ser Leu Pro Pro770 775 780Glu Asp Val Arg Ile Arg Met Leu Asn Leu Thr Thr Leu Arg Ile Ser785 790 795 800Trp Lys Ala Pro Lys Ala Asp Gly Ile Asn Gly Ile Leu Lys Gly Phe805 810 815Gln Ile Val Ile Val Gly Gln Ala Pro Asn Asn Asn Arg Asn Ile Thr820 825 830Thr Asn Glu Arg Ala Ala Ser Val Thr Leu Phe His Leu Val Thr Gly835 840 845Met Thr Tyr Lys Ile Arg Val Ala Ala Arg Ser Asn Gly Gly Val Gly850 855 860Val Ser His Gly Thr Ser Glu Val Ile Met Asn Gln Asp Thr Leu Glu865 870 875 880Lys His Leu Ala Ala Gln Gln Glu Asn Glu Ser Phe Leu Tyr Gly Leu885 890 895Ile Asn Lys Ser His Val Pro Val Ile Val Ile Val Ala Ile Leu Ile900 905 910Ile Phe Val Val Ile Ile Ile Ala Tyr Cys Tyr Trp Arg Asn Ser Arg915 920 925Asn Ser Asp Gly Lys Asp Arg Ser Phe Ile Lys Ile Asn Asp Gly Ser930 935 940Val His Met Ala Ser Asn Asn Leu Trp Asp Val Ala Gln Asn Pro Asn945 950 955 960Gln Asn Pro Met Tyr Asn Thr Ala Gly Arg Met Thr Met Asn Asn Arg965 970 975Asn Gly Gln Ala Leu Tyr Ser Leu Thr Pro Asn Ala Gln Asp Phe Phe980 985 990Asn Asn Cys Asp Asp Tyr Ser Gly Thr Met His Arg Pro Gly Ser Glu995 1000 1005His His Tyr His Tyr Ala Gln Leu Thr Gly Gly Pro Gly Asn Ala Met1010 1015 1020Ser Thr Phe Tyr Gly Asn Gln Tyr His Asp Asp Pro Ser Pro Tyr Ala1025 1030 1035 1040Thr Thr Thr Leu Val Leu Ser Asn Gln Gln Pro Ala Trp Leu Asn Asp1045 1050 1055Lys Met Leu Arg Ala Pro Ala Met Pro Thr Asn Pro Val Pro Pro Glu1060 1065 1070Pro Pro Ala Arg Tyr Ala Asp His Thr Ala Gly Arg Arg Ser Arg Ser1075 1080 1085Ser Arg Ala Ser Asp Gly Arg Gly Thr Leu Asn Gly Gly Leu His His1090 1095 1100Arg Thr Ser Gly Ser Gln Arg Ser Asp Ser Pro Pro His Thr Asp Val1105 1110 1115 1120Ser Tyr Val Gln Leu His Ser Ser Asp Gly Thr Gly Ser Ser Lys Glu1125 1130 1135Arg Thr Gly Glu Arg Arg Thr Pro Pro Asn Lys Thr Leu Met Asp Phe1140 1145 1150Ile Pro Pro Pro Pro Ser Asn Pro Pro Pro Pro Gly Gly His Val Tyr1155 1160 1165Asp Thr Ala Thr Arg Arg Gln Leu Asn Arg Gly Ser Thr Pro Arg Glu1170 1175 1180Asp Thr Tyr Asp Ser Val Ser Asp Gly Ala Phe Ala Arg Val Asp Val1185 1190 1195 1200Asn Ala Arg Pro Thr Ser Arg Asn Arg Asn Leu Gly Gly Arg Pro Leu1205 1210 1215Lys Gly Lys Arg Asp Asp Asp Ser Gln Arg Ser Ser Leu Met Met Asp1220 1225 1230Asp Asp Gly Gly Ser Ser Glu Ala Asp Gly Glu Asn Ser Glu Gly Asp1235 1240 1245Val Pro Arg Gly Gly Val Arg Lys Ala Val Pro Arg Met Gly Ile Ser1250 1255 1260Ala Ser Thr Leu Ala His Ser Cys Tyr Gly Thr Asn Gly Thr Ala Gln1265 1270 1275 1280Arg Phe Arg Ser Ile Pro Arg Asn Asn Gly Ile Val Thr Gln Glu Gln1285 1290 1295Thr181651PRThuman 18Met Lys Trp Lys His Val Pro Phe Leu Val Met Ile Ser Leu Leu Ser1 5 10 15Leu Ser Pro Asn His Leu Phe Leu Ala Gln Leu Ile Pro Asp Pro Glu20 25 30Asp Val Glu Arg Gly Asn Asp His Gly Thr Pro Ile Pro Thr Ser Asp35 40 45Asn Asp Asp Asn Ser Leu Gly Tyr Thr Gly Ser Arg Leu Arg Gln Glu50 55 60Asp Phe Pro Pro Arg Ile Val Glu His Pro Ser Asp Leu Ile Val Ser65 70 75 80Lys Gly Glu Pro Ala Thr Leu Asn Cys Lys Ala Glu Gly Arg Pro Thr85 90 95Pro Thr Ile Glu Trp Tyr Lys Gly Gly Glu Arg Val Glu Thr Asp Lys100 105 110Asp Asp Pro Arg Ser His Arg Met Leu Leu Pro Ser Gly Ser Leu Phe115 120 125Phe Leu Arg Ile Val His Gly Arg Lys Ser Arg Pro Asp Glu Gly Val130 135 140Tyr Val Cys Val Ala Arg Asn Tyr Leu Gly Glu Ala Val Ser His Asn145 150 155 160Ala Ser Leu Glu Val Ala Ile Leu Arg Asp Asp Phe Arg Gln Asn Pro165 170 175Ser Asp Val Met Val Ala Val Gly Glu Pro Ala Val Met Glu Cys Gln180 185 190Pro Pro Arg Gly His Pro Glu Pro Thr Ile Ser Trp Lys Lys Asp Gly195 200 205Ser Pro Leu Asp Asp Lys Asp Glu Arg Ile Thr Ile Arg Gly Gly Lys210 215 220Leu Met Ile Thr Tyr Thr Arg Lys Ser Asp Ala Gly Lys Tyr Val Cys225 230 235 240Val Gly Thr Asn Met Val Gly Glu Arg Glu Ser Glu Val Ala Glu Leu245 250 255Thr Val Leu Glu Arg Pro Ser Phe Val Lys Arg Pro Ser Asn Leu Ala260 265 270Val Thr Val Asp Asp Ser Ala Glu Phe Lys Cys Glu Ala Arg Gly Asp275 280 285Pro Val Pro Thr Val Arg Trp Arg Lys Asp Asp Gly Glu Leu Pro Lys290 295 300Ser Arg Tyr Glu Ile Arg Asp Asp His Thr Leu Lys Ile Arg Lys Val305 310 315 320Thr Ala Gly Asp Met Gly Ser Tyr Thr Cys Val Ala Glu Asn Met Val325 330 335Gly Lys Ala Glu Ala Ser Ala Thr Leu Thr Val Gln Glu Pro Pro His340 345 350Phe Val Val Lys Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val355 360 365Thr Phe Gln Cys Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp370 375 380Arg Arg Glu Gly Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln385 390 395 400Ser Ser Ser Arg Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr405 410 415Asn Val Gln Arg Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn420 425 430Val Ala Gly Ser Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val435 440 445Ile Ala Asp Arg Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln450 455 460Thr Val Ala Val Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly465 470 475 480Ser Pro Val Pro Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser485 490 495Thr Gln Asp Ser Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile500 505 510Arg Tyr Ala Lys Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser515 520 525Thr Pro Ser Gly Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu530 535 540Phe Gly Val Pro Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile545 550 555 560Pro Ser Ala Pro Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr565 570 575Val Thr Leu Ser Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr580 585 590Ser Tyr Ile Ile Glu Ala Phe Ser His Ala Ser Gly Ser Ser Trp Gln595 600 605Thr Val Ala Glu Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu610 615 620Lys Pro Asn Ala Ile Tyr Leu Phe Leu Val Arg Ala Ala Asn Ala Tyr625 630 635 640Gly Ile Ser Asp Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp645 650 655Val Leu Pro Thr Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu660 665 670Leu Gly Asn Ala Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser675 680 685Ser Ser Ile Glu Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile690 695 700Gln Gly Tyr Lys Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu705 710 715 720Ser Asp Trp Leu Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val725 730 735Val Ile Pro Asp Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg740 745 750Pro Phe Phe Asn Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala755 760 765Lys Thr Leu Glu Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val770 775 780Ser Lys Asn Asp Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro785 790 795 800Pro Pro Glu Asp Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp805 810 815Cys Leu Gly Asn Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly820 825 830Ser Thr Phe Ser Val Val Ile Pro Phe Leu Val Pro Gly Ile Arg Tyr835 840 845Ser Val Glu Val Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser850 855 860Glu Pro Gln Phe Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro865 870 875 880Glu Asp Gln Val Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln885 890 895Pro Ala Phe Ile Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met900 905 910Val Phe Ser Ile Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu915 920 925Thr Ser Thr Tyr Ala Gly Ile Arg Lys Val Pro Ser Phe Thr Phe Thr930 935 940Pro Thr Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser Ser Gly Gly945 950 955 960Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln Pro Trp Leu965 970 975Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp Cys Ser Ile980 985 990Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn Leu Thr Thr995 1000 1005Tyr Ser Arg Pro Ala Asp Cys Ile Ala Asn Tyr Asn Asn Gln Leu Asp1010 1015 1020Asn Lys Gln Thr Asn Leu Met Leu Pro Glu Ser Thr Val Tyr Gly Asp1025 1030 1035 1040Val Asp Leu Ser Asn Lys Ile Asn Glu Met Lys Thr Phe Asn Ser Pro1045 1050 1055Asn Leu Lys Asp Gly Arg Phe Val Asn Pro Ser Gly Gln Pro Thr Pro1060 1065 1070Tyr Ala Thr Thr Gln Leu Ile Gln Ser Asn Leu Ser Asn Asn Met Asn1075 1080 1085Asn Gly Ser Gly Asp Ser Gly Glu Lys His Trp Lys Pro Leu Gly Gln1090 1095 1100Gln Lys Gln Glu Val Ala Pro Val Gln Tyr Asn Ile Val Glu Gln Asn1105 1110 1115 1120Lys Leu Asn Lys Asp Tyr Arg Ala Asn Asp Thr Val Pro Pro Thr Ile1125 1130 1135Pro Tyr Asn Gln Ser Tyr Asp Gln Asn Thr Gly Gly Ser Tyr Asn Ser1140 1145 1150Ser Asp Arg Gly Ser Ser Thr Ser Gly Ser Gln Gly His Lys Lys Gly1155 1160 1165Ala Arg Thr Pro Lys Val Pro Lys Gln Gly Gly Met Asn Trp Ala Asp1170 1175 1180Leu Leu Pro Pro Pro Pro Ala His Pro Pro Pro His Ser Asn Ser Glu1185 1190 1195 1200Glu Tyr Asn Ile Ser Val Asp Glu Ser Tyr Asp Gln Glu Met Pro Cys1205 1210 1215Pro Val Pro Pro Ala Arg Met Tyr Leu Gln Gln Asp Glu Leu Glu Glu1220 1225 1230Glu Glu Asp Glu Arg Gly Pro Thr Pro Pro Val Arg Gly Ala Ala Ser1235 1240 1245Ser Pro Ala Ala Val Ser Tyr Ser His Gln Ser Thr Ala Thr Leu Thr1250 1255 1260Pro Ser Pro Gln Glu Glu Leu Gln Pro Met Leu Gln Asp Cys Pro Glu1265 1270 1275 1280Glu Thr Gly His Met Gln His Gln Pro Asp Arg Arg Arg Gln Pro Val1285 1290 1295Ser Pro Pro Pro Pro Pro Arg Pro Ile Ser Pro Pro His Thr Tyr Gly1300 1305 1310Tyr Ile Ser Gly Pro Leu Val Ser Asp Met Asp Thr Asp Ala Pro Glu1315 1320 1325Glu Glu Glu Asp Glu Ala Asp Met Glu Val Ala Lys Met Gln Thr Arg1330 1335 1340Arg Leu Leu Leu Arg Gly Leu Glu Gln Thr Pro Ala Ser Ser Val Gly1345 1350 1355 1360Asp Leu Glu Ser Ser Val Thr Gly Ser Met Ile Asn Gly Trp Gly Ser1365 1370 1375Ala Ser Glu Glu Asp Asn Ile Ser Ser Gly Arg Ser Ser Val Ser Ser1380 1385 1390Ser Asp Gly Ser Phe Phe Thr Asp Ala Asp Phe Ala Gln Ala Val

Ala1395 1400 1405Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala Arg Arg Gln Met Gln1410 1415 1420Asp Ala Ala Gly Arg Arg His Phe His Ala Ser Gln Cys Pro Arg Pro1425 1430 1435 1440Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser Ala Ala Val Met Gln1445 1450 1455Lys Thr Arg Pro Ala Lys Lys Leu Lys His Gln Pro Gly His Leu Arg1460 1465 1470Arg Glu Thr Tyr Thr Asp Asp Leu Pro Pro Pro Pro Val Pro Pro Pro1475 1480 1485Ala Ile Lys Ser Pro Thr Ala Gln Ser Lys Thr Gln Leu Glu Val Arg1490 1495 1500Pro Val Val Val Pro Lys Leu Pro Ser Met Asp Ala Arg Thr Asp Arg1505 1510 1515 1520Ser Ser Asp Arg Lys Gly Ser Ser Tyr Lys Gly Arg Glu Val Leu Asp1525 1530 1535Gly Arg Gln Val Val Asp Met Arg Thr Asn Pro Gly Asp Pro Arg Glu1540 1545 1550Ala Gln Glu Gln Gln Asn Asp Gly Lys Gly Arg Gly Asn Lys Ala Ala1555 1560 1565Lys Arg Asp Leu Pro Pro Ala Lys Thr His Leu Ile Gln Glu Asp Ile1570 1575 1580Leu Pro Tyr Cys Arg Pro Thr Phe Pro Thr Ser Asn Asn Pro Arg Asp1585 1590 1595 1600Pro Ser Ser Ser Ser Ser Met Ser Ser Arg Gly Ser Gly Ser Arg Gln1605 1610 1615Arg Glu Gln Ala Asn Val Gly Arg Arg Asn Ile Ala Glu Met Gln Val1620 1625 1630Leu Gly Gly Tyr Glu Arg Gly Glu Asp Asn Asn Glu Glu Leu Glu Glu1635 1640 1645Thr Glu Ser165019434PRThumanmisc_feature(285)..(396)note="Xaa signifies gap in sequence" 19Gln Ile Val Ala Gln Gly Arg Thr Val Thr Phe Pro Cys Glu Thr Lys1 5 10 15Gly Asn Pro Gln Pro Ala Val Phe Trp Gln Lys Glu Gly Ser Gln Asn20 25 30Leu Leu Phe Pro Asn Gln Pro Gln Gln Pro Asn Ser Arg Cys Ser Val35 40 45Ser Pro Thr Gly Asp Leu Thr Ile Thr Asn Ile Gln Arg Ser Asp Ala50 55 60Gly Tyr Tyr Ile Cys Gln Ala Leu Thr Val Ala Gly Ser Ile Leu Ala65 70 75 80Lys Ala Gln Leu Glu Val Thr Asp Val Leu Thr Asp Arg Pro Pro Pro85 90 95Ile Ile Leu Gln Gly Pro Ala Asn Gln Thr Leu Ala Val Asp Gly Thr100 105 110Ala Leu Leu Lys Cys Lys Ala Thr Gly Asp Pro Leu Pro Val Ile Ser115 120 125Trp Leu Lys Glu Gly Phe Thr Phe Pro Gly Arg Asp Pro Arg Ala Thr130 135 140Ile Gln Glu Gln Gly Thr Leu Gln Ile Lys Asn Leu Arg Ile Ser Asp145 150 155 160Thr Gly Thr Tyr Thr Cys Val Ala Thr Ser Ser Ser Gly Glu Ala Ser165 170 175Trp Ser Ala Val Leu Asp Val Thr Glu Ser Gly Ala Thr Ile Ser Lys180 185 190Asn Tyr Asp Leu Ser Asp Leu Pro Gly Pro Pro Ser Lys Pro Gln Val195 200 205Thr Asp Val Thr Lys Asn Ser Val Thr Leu Ser Trp Gln Pro Gly Thr210 215 220Pro Gly Thr Leu Pro Ala Ser Ala Tyr Ile Ile Glu Ala Phe Ser Gln225 230 235 240Ser Val Ser Asn Ser Trp Gln Thr Val Ala Asn His Val Lys Thr Thr245 250 255Leu Tyr Thr Val Arg Gly Leu Arg Pro Asn Thr Ile Tyr Leu Phe Met260 265 270Val Arg Ala Ile Asn Pro Lys Val Ser Val Thr Gln Xaa Lys Pro Gln275 280 285Lys Asn Asn Gly Ser Thr Trp Ala Asn Val Pro Leu Pro Pro Pro Pro290 295 300Val Gln Pro Leu Pro Gly Thr Glu Leu Glu His Tyr Ala Val Glu Gln305 310 315 320Gln Glu Asn Gly Tyr Asp Ser Asp Ser Trp Cys Pro Pro Leu Pro Val325 330 335Gln Thr Tyr Leu His Gln Gly Leu Glu Asp Glu Leu Glu Glu Asp Asp340 345 350Asp Arg Val Pro Thr Pro Pro Val Arg Gly Val Ala Ser Ser Pro Ala355 360 365Ile Ser Phe Gly Gln Gln Ser Thr Ala Thr Leu Thr Pro Ser Pro Arg370 375 380Glu Glu Met Gln Pro Met Leu Gln Ala Ser Pro Xaa Phe Thr Ser Ser385 390 395 400Gln Arg Pro Arg Pro Thr Ser Pro Phe Ser Thr Asp Ser Asn Thr Ser405 410 415Ala Ala Leu Ser Gln Ser Gln Arg Pro Arg Pro Thr Lys Lys His Lys420 425 430Gly Gly20148PRTmouse 20Ala Gln Ala Val Ala Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala1 5 10 15Arg Arg Gln Met Gln Asp Ala Ala Gly Arg Arg His Phe His Ala Ser20 25 30Gln Cys Pro Arg Pro Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser35 40 45Ala Val Val Ile Gln Lys Ala Arg Pro Ala Lys Lys Gln Lys His Gln50 55 60Pro Gly His Leu Arg Arg Glu Ala Tyr Ala Asp Asp Leu Pro Pro Pro65 70 75 80Pro Val Pro Pro Pro Ala Ile Lys Ser Pro Thr Val Gln Ser Lys Ala85 90 95Gln Leu Glu Val Arg Pro Val Met Val Pro Lys Leu Ala Ser Ile Glu100 105 110Ala Arg Thr Asp Arg Ser Ser Asp Arg Lys Gly Gly Ser Tyr Lys Gly115 120 125Arg Glu Ala Leu Asp Gly Arg Gln Val Thr Asp Leu Arg Thr Asn Pro130 135 140Ser Asp Pro Arg145

Claims

1. An isolated Slit polypeptide comprising a vertebrate species-specific Slit fragment.

2. A method of modulating the interaction of Robo and a Robo ligand, or said method comprising a step ofcombining a Robo polypeptide, a Slit polypeptide according to claim 1, and a modulator under conditions whereby, but for the presence of the modulator, the Robo and Slit polypeptide engage in a first interaction, wherein the Slit polypeptide specifically binds, activates or inhibits the activation of the Robo polypeptide andwhereby the Robo and Slit polypeptide engage in a second interaction different from the first interaction.

3. A method of identifying agents which modulate the interaction of Robo and a Robo ligand, said method comprising the method of claim 2, wherein the steps are:combining a Robo polypeptide, a Slit polypeptide and a candidate agent under conditions whereby, but for the presence of the agent, the Robo and Slit polypeptides engage in a first interaction, wherein the Slit polypeptide specifically binds, activates or inhibits the activation of the Robo polypeptide anddetermining a second interaction of the Robo and Slit polypeptides in the presence of the agent,wherein a difference between the first and second interactions indicates that the agent modulates the interaction of the Robo and Slit polypeptides

4. A method according to claim 3, wherein the modulator is a dominant negative form of the Robo or Slit polypeptide.

5. An isolated vertebrate Slit polypeptide according to claim 1, wherein said vertebrate is human, mouse or rat.

6. A recombinant nucleic acid encoding a vertebrate Slit polypeptide according to claim 5.

7. A recombinant nucleic acid according to claim 6 and comprising a strand of SEQ ID NO:01, or a fragment thereof having at least 24 consecutive nucleotides thereof, and sufficient to specifically hybridize with a polynucleotide having the sequence defined by the corresponding opposite strand of SEQ ID No:01, and is other than a natural drosophila Slit sequence.

8. A method for specifically detecting a vertebrate Slit protein, comprising the steps of:specifically binding an antibody according to claim 8 to the Slit protein; andspecifically detecting a resultant specific binding as an indication of the presence of the Slit protein.

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