METHODS OF IDENTIFYING GLYCOPEPTIDES RECOGNIZED BY DISEASE-ASSOCIATED AUTO-ANTIBODIES

Abstract

Methods for identifying glycopeptides and more particularly glycopeptide epitopes that are specifically recognized by disease-associated auto-antibodies are provided. In some aspects the auto-antibodies are cancer-associated or autoimmune disease associated. In other aspects, methods of diagnosing a patient with cancer or an autoimmune disease, or for eliciting an immune response in a mammalian host directed to the glycopeptides of the invention are provided.

Description

CROSS REFERENCE TO PRIOR APPLICATION

[0001] This application is a continuation of U.S. application Ser. No. 11/987,034, filed Jan. 7, 2011, which claims priority to U.S. Provisional Application Ser. No. 61/293,583, filed Jan. 8, 2010 and Ser. No 61/294,477 filed Jan. 12, 2010, all of which are incorporated by reference herein in their entireties.

FIELD OF THE INVENTION

[0002] The present invention is related to methods for identifying glycopeptides and more particularly glycopeptide epitopes that are specifically recognized by disease-associated auto-antibodies. In some aspects the auto-antibodies are cancer-associated or autoimmune disease-associated. The invention also relates to methods of diagnosing a patient with cancer or an autoimmune disease, or for eliciting an immune response in a mammalian host directed to a glycopeptide of the invention. The invention also relates to novel individual glycopeptides as well as to panels of glycopeptides (whether individually novel or not) useful for autoantibody detection purposes.

BACKGROUND OF INVENTION

[0003] Malignant transformation of cells is virtually always accompanied by alterations in the posttranslational modifications (PTMs) of proteins, and one of the best documented examples is the abundant mucin-type O-glycosylation (hereafter referred to as O-glycosylation) found on mucins and other O-glycoproteins (Tarp and Clausen 2008). Tumor-associated changes in expression of O-glycoproteins and/or in their aberrant glycosylation, create a diverse set of unusual molecular structures found on the surface of cancer cells as well as in secretions. These molecular structures generally represent glycoproteins with truncated immature O-glycans, to which the immune system of man is not normally exposed except, in some cases, as biosynthetic intermediates and then only in the secretory pathway. Therefore, these structures may represent a different form of tumor-associated antigens (one not necessarily based on differential expression level or sequence mutation), to which individuals lack immunological tolerance and thus provoke both auto-antibodies and cell mediated immunity (Anderton 2004; Doyle and Mamula 2005; Doyle and Mamula 2001).

[0004] Several mouse antibodies have been isolated and characterized as having unique binding specificity for combined glycopeptide epitopes that include both the peptide sequence as well as the aberrant PTM (hereinafter "APTM") for efficient binding. Examples include monoclonal antibodies that specifically recognize distinct O-glycopeptides from MUC2, MUC1, and other glycoproteins (Reis et al. 1998; Sorensen et al. 2006; Danielczyk et al. 2006; Dian et al. 2009; Li et al. 2009; Takeuchi et al. 2002; Clark et al. 1998).

[0005] There exist examples of human antibodies with selective or specific reactivity with PTM-modified proteins, but these are generally limited to inflammatory and autoimmune diseases (Anderton 2004; Doyle and Mamula 2005; Doyle and Mamula 2001). Furthermore, human hydridoma technologies have identified natural IgM antibodies that react with glycoforms of proteins but the nature of the epitopes has not been fully clarified (Rauschert et al. 2008; Vollmers and Brandlein 2009; Brandlein et al. 2004a; Brandlein et al. 2004b; Rasmussen and Ditzel 2009). A major drawback of current technologies to screen for auto-antibodies directed against APTM proteins is the lack of high through-put methods for identifying antigens, more particularly epitopes, for generating pertinent antigens and for screening such antigens.

[0006] In principle, cancer-associated auto-antibodies represent appealing potential biomarkers. Auto-antibodies may develop early in carcinogenesis, at the time tumor-associated antigens appear on premalignant or malignant lesions. Antibody responses can produce relatively high concentrations in circulation with a long circulation time, and they can be detected with sensitive and specific methods (see, (Lu et al. 2008; Anderson and Labaer 2005)). In contrast, antigens produced by small premalignant or malignant lesions are generally produced in vanishingly small levels that due to dilution and clearance from blood may not be detectable by conventional techniques. Discovery and characterization of specific auto-antibodies to cancer antigens have been undertaken using different approaches in the past. Classical studies identified such antibodies reactive with tumor cells, tissues, or isolated proteins (Kawabata et al. 2007), but distinct molecular features of binding epitopes have generally not resulted from these approaches.

[0007] More recent proteome-wide screening techniques have included expressed cDNA libraries (SEREX) (Sahin et al. 1995), protein and peptide arrays (Stockert et al. 1998; Pereira-Faca et al. 2007), random or designed phage displays (Mintz et al. 2003), and more recently self-assembling protein arrays (Ramachandran et al. 2008; Anderson et al. 2008). Cancer-associated auto-antibodies characterized to date have been found to bind intracellular proteins with functions important in cell cycle regulation, such as GPR78 (Mintz et al. 2003), p53 (Lubin et al. 1993), NY-ESO-1, and CDC25 (Liu et al. 2008), but also some cell membrane glycoproteins such as MUC1 (Snijdewint et al. 1999), HER2 (Chapman et al. 2007) and Mesothelin (Hellstrom et al. 2008).

[0008] Auto-antibodies are believed to be induced as a result of altered expression of proteins and altered molecular structure due to mutations, alternative splicing and post-translational events such as protein processing and aberrant enzymatic modifications including glycosylation (Anderton 2004; Doyle and Mamula 2005; Doyle and Mamula 2001). These events induce breakage of tolerance and immunity may result. Surprisingly, however, few disease- or more specifically cancer-associated auto-antibody epitopes have been identified and molecularly defined despite considerable efforts and broad proteome screening. This is due to limitations in methods for identification of such auto-antibodies, in that, before the present invention, the appropriate antigen epitopes have not been determined and hence not tested to lead to identification of disease-associated antibodies, to serve as substrates for the detection of disease or to serve as prototype vaccines for induction of immunity against the epitopes of these autoantibodies.

[0009] There are few known examples of disease-associated human antibodies to proteins involving glycosylation. One important example is an immunodominant epitope in type II collagen comprising a glycosylated hydroxylysine residue that is involved in collagen-induced arthritis (Backlund et al. 2002). Glycosylation may also modulate protein processing and hence affect exposure of new epitopes as shown in Rasmussen's encephalitis, where an N-glycan blocks proteolysis of a neuronal glutamate receptor and a short preceding peptide epitope (Gahring et al. 2001). Several human monoclonal antibodies have been shown to be directed to epitopes affected by glycosylation (Rauschert et al. 2008; Vollmers and Brandlein 2009; Brandlein et al. 2004a; Brandlein et al. 2004b; Rasmussen and Ditzel 2009), but the nature of the molecular epitopes remains undefined.

[0010] There is therefore a need to develop methods for the identification of PTM-containing peptides, such as aberrant glycopeptides (hereinafter "AGP"), that are specifically recognized by disease-associated auto-antibodies. The present invention provides such methods. There is also a need for improved diagnostic tools, such as AGP, that would permit early detection of disease, notably cancer, for example by being used as substrates to capture disease-associated autoantibodies.

SUMMARY OF INVENTION

[0011] In one embodiment, the invention provides a method for identifying glycopeptides reactive with cancer-associated auto-antibodies, the method comprising: providing a panel comprising peptides, at least a plurality of the peptides comprising one or more sites amenable to glycosylation, wherein at least one of the glycosylation sites has been modified with a glycan to form a glycopeptide having a peptide portion and a glycan portion; contacting the panel with an antibody-containing sample from a patient with cancer; and identifying glycopeptides in the panel that (i) are selectively recognized by antibodies in the sample, but not by antibodies in a control sample and (ii) are recognized by such antibodies in the sample through recognition of both the peptide portion and the glycan portion and not through recognition of either the peptide or the glycan alone.

[0012] In another embodiment, a panel of the invention comprises a plurality of peptides having an amino acid sequence comprising at least one serine or threonine residue, wherein the residue is a glycosylation site. In yet another embodiment, the at least one serine or threonine residue is at about the middle of the amino acid sequence. In still another embodiment, a panel of the invention comprises mutants of the peptides. The number of peptides in a panel may vary and may be at least 8 peptides, for example between about 8 and about 30 peptides, such as 10, 12, 15 or 20 peptides. A number of peptides in excess of 30 is also within the invention. The upper limit of peptides in a panel is limited by practical considerations (e.g., how many peptides can fit on a substrate) or cost-benefit considerations. Preferably the peptides are selected from the group consisting of SEQ ID NOs 15, 36, 49, and 82-146.

[0013] In a specific aspect of the above embodiment, providing a method for identifying glycopeptides reactive with cancer-associated auto-antibodies, the method further comprises identifying cancer-associated glycopeptide epitopes. In another aspect, the method further comprising elucidating the epitope structure of the identified glycopeptide epitopes.

[0014] In a specific embodiment, the invention provides a method for determining whether a patient has cancer, the method comprising: contacting an antibody-containing sample from the patient with a panel comprising peptides at least a plurality of which are glycopeptides, each glycopeptide comprising a glycopeptide epitope, the epitope having been previously determined (i) to be selectively recognized by a subset of antibodies in sera from cancer patients, which subset recognizes neither (a) the corresponding naked peptides of the panel when not glycosylated; nor (b) the corresponding glycan when not bound to the peptide; and (ii) not to be recognized by antibodies in control sera; contacting the panel with an antibody-containing sample from the patient; determining if antibodies in the sample are bound to glycopeptides of the panel; and concluding either that the patient has cancer if the sample comprises antibodies that bind to at least one of the glycopeptides in the panel; or that the patient does not have cancer if the sample does not comprise antibodies that bind to at least one glycopeptide in the panel.

[0015] In one aspect of the above embodiment providing a method for determining whether a patient has cancer, the patient is newly diagnosed with cancer. In another aspect, the method further comprises concluding either that the patient has cancer if the sample comprises IgG antibodies that bind to at least one of the glycopeptides in the panel; or that the patient does not have cancer if the sample does not comprise IgG antibodies that bind to at least one glycopeptide in the panel. In yet another aspect, the patient is diagnosed with cancer if said sample comprises antibodies that specifically recognize one or more of the glycopeptides having an amino acid sequence selected from the group consisting of: SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0016] In another embodiment, the invention provides a method for eliciting an immune response in a patient, the method comprising administering to the patient a composition comprising (i) a glycopeptide consisting essentially of an amino acid sequence selected from the group consisting of SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146), in an effective amount for eliciting the immune response; and (ii) a suitable adjuvant.

[0017] In a specific aspect of the above embodiment, providing a method for eliciting an immune response in a patient, the immune response comprises an IgG antibody response. In another aspect, the immune response is an anti-cancer immune response. In yet another aspect, the cancer is selected from the group consisting of breast cancer; colon cancer; ovarian cancer; cervical cancer; pancreatic cancer; prostatic cancer; liver cancer; kidney cancer; brain cancer; hematological cancer; testis cancer; head and neck cancer; and lung cancer.

[0018] In one aspect of the above embodiment, providing a method for eliciting an immune response in a patient, the glycopeptide is modified with an O-glycan at at least one amino acid residue. In another aspect, the at least one amino acid residue is a serine or threonine residue.

[0019] In any of the above embodiments of the invention, each of the peptides in the panel may be about 2 to about 50 amino acid residues in length. In other aspects, each of the peptides in the panel may be about 4 to about 25 amino acid residues in length.

[0020] In certain aspects of the above embodiments of the invention, the amino acid sequence of the peptides of the panel is a fragment found in a protein or variant of said protein or a conservative mutant. In some of the above embodiments, the identified glycopeptides of the panel are found in at least one glycoprotein that is aberrantly glycosylated in cancer cells. In yet other of the above embodiments, glycopeptides identified by the methods of the present invention are found in at least one glycoprotein that is overexpressed in cancer cells.

[0021] In other aspects, the glycopeptides are synthesized synthetically or chemoenzymatically. In still other of the above aspects, the glycopeptides of the panel are partially glycosylated peptides when immobilized on the panel.

[0022] In certain of the above embodiments, the control sample contains pooled samples from a plurality of control individuals.

[0023] In certain of the above aspects of the invention, the panel is a microarray slide. In other aspects, the glycopeptides identified by the methods of the invention are selectively recognized by an IgG antibody. In certain of the above embodiments, the panel comprises one or more glycopeptides having an amino acid sequence selected from the group consisting of: SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0024] In another aspect, the invention provides a glycopeptide comprising an amino acid sequence selected from the group consisting of SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0025] In yet another aspect, the invention provides a glycopeptide consisting essentially of an amino acid sequence selected from the group consisting of SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0026] In still another aspect, the invention provides a glycopeptide consisting of an amino acid sequence selected from the group consisting of SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49), LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0027] In another aspect, the invention provides a pharmaceutical composition comprising one or more glycopeptides selected from the group consisting of SEQ ID NOs 15, 36, 49, and 82-146. Preferably, the pharmaceutical composition comprises one or more glycopeptides selected from the group consisting of LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0028] In yet another aspect, the invention provides for a panel of glycopeptides comprising at least a plurality of glycopeptides, each glycopeptide comprising a glycopeptide epitope, said epitope having been previously determined (i) to be selectively recognized by a subset of antibodies in sera from cancer patients, which subset recognizes neither (a) the corresponding naked peptides of said panel when not glycosylated; nor (b) the corresponding glycan when not bound to said peptide; and (ii) not to be recognized by antibodies in control sera, said plurality comprising at least 8 glycopeptides selected from the group consisting of glycopeptides having SEQ IDs 15, 36, 49, and 82-146. Preferably, the plurality of glycopeptides comprises SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

[0029] In any of the above embodiments of the invention, the glycopeptides of the panel may be glycosylated in situ, in solution, or in vivo by recombinant expression in a host cell. In any of the above embodiments, the glycopeptides of the panel may be treated with one or more exoglycosidases to expose O-glycans. In still other of the above embodiments, the glycan may be an O-glycan. In any of the above embodiments, the O-glycan may be a member selected from the group consisting of: Tn, STn, T, Truncated C3, Truncated C2, Truncated C4, non-capped type1-C3, non-capped type2-C2, non-capped type2-C4, GalNAca-Tn, SA-type1-C3, SLea-C3, LacDiNAc-C3, LacDiNAc-C2, and LacDiNAc-C4.

[0030] In certain of the above embodiments, at least one glycopeptide in the panel is not a glycopeptide comprising a glycosylated GSTA motif.

[0031] In certain of the above embodiments, none of the glycopeptides in the panel are glycopeptides comprising a glycosylated GSTA motif.

[0032] These and other aspects of the present invention will be apparent to those of ordinary skill in the art in light of the present specification, claims and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIG. 1 is an image of a microarray slide and illustrates an example of on-slide (in vitro) glycosylation with a polypeptide GalNAc-transferase, GalNAc-T3, to glycosylate peptides and GalNAc glycopeptides with additional unsubstituted Ser/Thr residues to enhance number of O-glycans. FIG. 1A: GalNAc-T3 untreated array, FIG. 1B: GalNAc-T3 treated array.

[0034] FIG. 2 is an image of a microarray slide and illustrates reactivities of serum (1:25 dilution) from a newly diagnosed prostate cancer patient (#762 in FIG. 2A) and a normal control serum from an individual that does not have prostate cancer (#174 in FIG. 2B) on the library of 96 paired peptides/GalNAc-glycopeptides as designated in Table II.

[0035] FIGS. 3-5 are bar graphs and illustrate microarray screening results from 147 cancer and 31 control sera on the glycopeptide pairs identified herein as #20/21 (FIG. 3), #62/63 (FIG. 4), or #86/87 (FIG. 5).

[0036] FIG. 6A-C are bar graphs showing 8 selected glycopeptides (i.e., 889, 275a, 585, 893, 931-C3, 852-C3, 873, and 690) reactive with IgG from newly diagnosed cancer patients compared to normal control sera. FIG. 6A: glycopeptides 889 (SEQ ID NO: 134), 275a (SEQ ID NO: 86), and 585 (SEQ ID NO: 109). FIG. 6B: glycopeptides 893 (SEQ ID NO: 135), 931-C3 (SEQ ID NO: 146), and 852-C3 (SEQ ID NO: 145). FIG. 6C: gycopeptides 873 (SEQ ID NO:132) and 690 (SEQ ID NO: 116). The following human sera were used for FIGS. 6A and 6B: 32 ovarian cancer (O), 38 breast cancer (B), 54 colon cancer (C), 17 lung cancer (L), as well as 145 normal sera (N) (Asterand Corp). The following sera were used for FIG. 6C: 32 ovarian cancer (O), 38 breast cancer (O), 54 colon cancer (C), 42 lung cancer (L), 52 pancreatic cancer (Pn), 35 prostatic cancer (Pr), 8 inflammatory disease (In), 8 benign controls (Bn), and 145 healthy controls (N). Sera were screened at 1:20 dilution. Anti-human-IgG conjugated to Cy3 was used as the secondary detection unit and slides were scanned with a GenePix 4200AL Microarray Scanner.

DETAILED DESCRIPTION OF THE INVENTION

[0037] The present invention provides methods for identification of peptides and peptide epitopes that comprise one or more posttranslational modification(s) (PTM(s)) ("PTM-peptides"/"PTM peptide epitopes") that are selectively recognized and bound by human disease-associated antibodies. These will include peptides that display an aberrant glycosylation pattern either due to an alteration in sequence (mutation) which invites a changed glycosylation pattern, or simply a glycosylation pattern not normally encountered in cancer-free individuals. Such peptides will constitute AGP as the term is introduced and hence defined above. For the removal of doubt, PTM-epitopes are not whole molecules; therefore, any discussion of such epitopes should be construed as applying to glycopeptides harboring such epitopes.

[0038] In some aspects, the PTM-peptides of the invention are useful for diagnosing a disease. In a specific embodiment, the present invention provides methods for diagnosing a patient with cancer. In another embodiment, the patient is newly diagnosed with cancer (i.e., diagnosed with cancer for the first time). In still other embodiments, a patient is diagnosed with an autoimmune disease.

[0039] In certain embodiments, the methods are useful for identifying PTM-peptides that are useful for eliciting in an individual an immune response directed against PTM-epitopes that are associated with disease, such as cancer. In certain embodiments, the immune response includes an auto-antibody response. In certain aspects, the induction of auto-antibodies to aberrant PTM-modified glycoproteins includes antibodies that specifically recognize O-glycopeptide epitopes.

[0040] As discussed, supra, malignant transformation of cells is virtually always accompanied by alterations in the posttranslational modifications (PTMs) of proteins, including O-glycosylation (Tarp and Clausen 2008). Tumor-associated changes in expression of O-glycoproteins and/or in their aberrant glycosylation, create a diverse set of unusual molecular structures found on the surface of cancer cells as well as in secretions. These molecular structures generally represent glycoproteins with truncated immature O-glycans, to which the immune system of man is not normally exposed except, in some cases, as biosynthetic intermediates and then only in the secretory pathway. Thus, one aspect of the PTM-peptides and PTM peptide epitopes identified by the methods of the present invention, i.e., AGP, is that they are not covered by immunological tolerance, and hence are potential targets for immunotherapeutic intervention. The aim of eliciting immune responses in a host using the AGP of the invention is the development of vaccines against the very disease that the AGP or other PTM-peptides are associated with.

[0041] Thus, AGP can cause induction of auto-antibodies to PTM-peptide epitopes. One aspect of this invention is the design and production of PTM-peptide libraries that represent or more accurately contain AGP associated with disease. In another aspect of the invention, high through-put methods for screening of disease-associated antibodies using such libraries are provided. In a specific embodiment, the invention relates to aberrant O-glycosylation of glycoproteins and more specifically, of glycopeptides. The present inventors believe one of the reasons why these disease-associated PTM-peptides and epitopes have eluded detection and especially association with disease is that the proteins bearing them in vivo are not or not necessarily mutated or overexpressed and hence would not be detected by ordinary, nontargeted, techniques.

[0042] PTM modifications can be the result of the addition of chemical groups to a protein, such as a phosphate group or a sugar moiety (e.g. acetylation of lysine and serine, glycosylation of asparagine, serine, threonine, hydroxyl-proline, lysine, methylation of arginine, histidine, and lysine, phosphorylation of serine, threonine, and tyrosine). They can also be the result of a conversion of an amino acid to a distinct structure, as in the deimination of arginine to citrulline or the deamidation of aspartic acid/asparagine to isoaspartic acid. While this invention is primarily concerned with glycosylation, it is evident that other aberrant posttranslational modifications as described herein may result in altered proteins harboring aberrant PTM-epitopes that may be the target of auto-immunity.

[0043] In some aspects of the present invention, the discriminating characteristic of the peptides and peptide epitopes useful in the present invention is that disease-associated antibodies bind selectively with the PTM-containing peptide and not with the same unmodified peptide or the PTM in the context of a different peptide sequence or another unrelated (e.g., artificial) carrier. In terms of AGP having diagnostic and/or immunogenic value, the disease-associated autoantibodies should recognize only the combination of the relevant glycosylated amino acid sequence and not the same glycan on a different peptide nor the same amino acid sequence bearing a different glycan. This will avoid false positives. It will be understood that the construction of peptide libraries and the availability of disease (e.g., cancer) and control sera that can be tested renders it unnecessary to know beforehand which AGP will be recognized by disease-associated antibodies in order to identify these antibodies. Once the disease-associated antibodies have been identified, they, or man-made versions thereof can be used to pinpoint the AGP that pulled relevant autoantibodies out of the sera.

[0044] In other aspects, the invention provides methods for the diagnosis of a patient with a disease, such as, e.g., cancer, wherein detection of antibodies in a sample from the patient that selectively recognize one or more combinations of disease-associated PTM-containing peptides is used to predict and diagnose disease. In still other embodiments, methods for treating a patient with cancer using compositions or vaccines comprising PTM-containing peptides or proteins of the invention are provided.

DEFINITIONS

[0045] As used herein, the term "immune response" includes an adaptive immune response, including a T cell response and B cell response. Thus, as used herein, the term "eliciting an immune response" means that an adaptive immune response is induced by administration of an appropriate immunogen (antigen or antigen plus carrier).

[0046] As used herein, an "anti-cancer immune response" is an immune response that is directed toward cancer cells or products secreted or shed from cancer cells. For example, an anti-cancer immune response may be characterized by tumor-specific antibodies and/or cytotoxic T cells that attack cancer cells or react specifically with a glycoprotein shed from cancer cells.

[0047] An "auto-antibody" is an antibody that specifically recognizes an epitope harbored by a self-product such as a protein, carbohydrate or lipid produced by healthy or diseased cells of an individual.

[0048] As used herein, the terms "selective binding" and "selective recognition" and their grammatical variants, of an epitope by an antibody means that the antibody binds with significantly greater affinity to the epitope compared to any other sequence or structure.

[0049] The term "epitope" refers to the part of an antigen that is specifically recognized and bound by an antibody. The term "glycopeptide epitope" is an epitope that includes both part of a peptide sequence and at least part of a glycan. The term "minimal epitope" or "minimal glycopeptide epitope" refers to the shortest glycopeptide that is recognized by antibodies recognizing the same epitope.

[0050] As used herein, the term "corresponding peptide" refers to the unglycosylated form of a glycopeptide of the invention. Thus, a glycopeptide and its corresponding peptide have the same amino acid sequence.

[0051] The term "glycan hapten" refers to glycan moiety independent of whether this is conjugated to a carrier, such as a peptide or protein (i.e. binding to the glycan hapten does not depend on the carrier such as protein or peptide to which the glycan is bound).

[0052] The terms "linear peptide" and "linear peptide epitope" means that the peptide or peptide epitope is non-conformational (i.e., antibody recognition and binding to the epitope does not depend upon the three-dimensional structure of the peptide).

[0053] The term "disease-associated antibody" means an antibody that is detected in a patient sample if the patient has the disease, but is not detected, or is present at significantly reduced levels compared to the patient sample, in a control sample. For example, a cancer-associated antibody is present in an antibody-containing sample from a patient with cancer, but is not present, or is present at significantly reduced levels in a sample from a patient without cancer. A useful distinguishing range of antibody levels is at least two or preferably three 3 fold higher levels in patients compared to healthy control. The term may also apply to groups of patients with a disease, where "disease-associated antibody" means an antibody that is detected with higher prevalence/incidence in the disease group compared to a healthy control group. A useful distinguishing prevalence (=specificity of assay) would be at least 70% and preferably 80 or 90%,

[0054] The term "aberrant glycosylation" means a glycoform that is not normally present on proteins expressed on the cell surface or secreted/shed from cells. An example of an aberrant glycoform is Tn (GalNAcα1-O-Ser/Thr), sialosyl-Tn (NeuAcα2-8GalNAcα1-O-Ser/Thr), and T (Galβ1-3GalNAcα1-O-Ser/Thr).

[0055] As used herein, a "glycosylatable peptide" or a peptide containing a "site amenable to glycosylation" refers to a peptide containing an amino acid residue that can be glycosylated. An example of a site amenable to glycosylation is a peptide site containing a serine or threonine amino acid residue, which can be O-glycosylated with N-acetyl-galactosamine (GalNAc).

[0056] As used herein, "variant" in addition to its understood meaning as a term of art includes any changes in a molecule from its wild-type form. For example, alleles, fragments, mutations, substitutions with natural or analog compounds, splice variants, glycosylations, species variants, and the like. The term is not limited to any one type of change or deviation from the wild type form or "normal" molecule. A "variant" also includes a polypeptide or enzyme which has at least 60% amino acid identity as determined by BLAST or FASTA algorithms, preferably at least 75%, most preferably at least 85%, and even more preferably at least 90%, and still more preferably at least 95%, and which has the same or substantially similar properties or functions as the native or parent protein or enzyme to which it is compared.

[0057] "Conservative mutants" are those in which a given amino acid residue in a protein or enzyme has been changed without altering the overall conformation and function of the polypeptide, including, but not limited to, replacement of an amino acid with one having similar properties (such as, for example, polarity, hydrogen bonding potential, acidic, basic, hydrophobic, aromatic, and the like). Amino acids with similar properties are well known in the art. For example, arginine, histidine and lysine are hydrophilic-basic amino acids and may be interchangeable. Similarly, isoleucine, a hydrophobic amino acid, may be replaced with leucine, methionine or valine. Such changes are expected to have little or no effect on the apparent molecular weight or isoelectric point of the protein or polypeptide.

[0058] An "organ-specific glycoprotein" is a glycoprotein which is only or which has substantial preferential expressed in a specific organ type. For example, the protein mucin 16 (MUC16) (SEQ ID NO: 57), is only expressed in epithelium of the female tract such as ovary and endometrium, and it is primarily overexpressed in tumors originating from these epithelia.

[0059] The term "clinical debut" refers to the first time a patient is diagnosed with a disease.

[0060] The term "immunogenic substitution" means the replacement of an amino acid residue with a different amino acid that causes the antigen to become immunogenic. For example, a substitution of an amino acid in a self protein, such as that which can result from a cancer mutation, or experimentally in a peptide, that renders the self protein immunogenic (i.e., breaks immunogenic tolerance to the self protein) is an immunogenic substitution. As another example, the substitution of a normally unmodified amino acid with a modified amino acid, wherein the modification itself (i.e., as a hapten, such as, e.g., Tn) is immunogenic, can render a protein immunogenic.

[0061] The term "subject" or "individual" as used herein refers to an animal having an immune system, preferably a mammal (e.g., rodent, such as mouse). In particular, the term encompasses humans.

[0062] As used herein, the term "about" or "approximately" usually means within an acceptable error range for the type of value and method of measurement. For example, it can mean within 20%, more preferably within 10%, and most preferably still within 5% of a given value or range. Alternatively, especially in biological systems, the term "about" means within about a log (i.e., an order of magnitude) preferably within a factor of two of a given value.

[0063] As used herein, the term "consisting essentially of" means that the glycopeptide sequence is not limited to a length of 20 amino acids, but can be longer or shorter. For example, a glycopeptide can be less than or more than 20 amino acids long as established during the course of optimizing the length of the peptide and efficient positioning of the glycosylated epitope that is specifically recognized by auto-antibodies in patients. Throughout the specification, the length of 20 amino acids is used as a convenient length and to ensure that the entire epitope is encompassed within the peptide. While most epitopes will be on the order of 5-10 amino acid residues, and hence contained within a 20 amino acid peptide, any size peptide is possible if sufficient to contain the epitope for antibody detection. In turn, longer peptides may allow for incorporation of additional epitopes which could provide different selectivity for cancer.

Post-Translational Modification of Peptides

[0064] In some aspects, the invention relates to the identification of disease-associated PTM-peptides, wherein the PTM-peptide is selectively recognized by disease-associated antibodies. Preferably, the disease-associated antibodies recognize neither the peptide alone nor the PTM alone or attached to a different peptide. In other words, the antibody epitope comprises both part of the protein backbone and part of the PTM.

[0065] The PTM may be one or more modifications of one or more residues of the amino acid sequence. The PTM can be any of the known PTMs involving enzymatic modifications of amino acids including glycosylation, phosphorylation, citrinylation, acetylation, methylation (Anderton 2004; Doyle and Mamula 2005; Doyle and Mamula 2001).

[0066] All 20 primary amino acids used by man are capable of undergoing some type of PTM. However, certain factors determine whether those modifications will take place. First, the location of the amino acid within the protein sequence affects both the type and frequency of modifications that may arise. Flanking residues can influence the conformation of the protein, potentially altering whether an enzyme has access to a certain amino acid or is exposed to a certain environment. The cellular location of the modifying enzyme, if required, will determine whether the modification occurs and disease-associated changes in localization of such enzymes may lead to aberrant posttranslational modifications. Previous modifications or proteolytic cleavages within a protein influence subsequent amino acid modifications within the same protein.

[0067] An exemplary PTM of the invention is mucin-type O-linked glycosylation ("O-glycosylation" or "O-linked glycosylation"). Examples of O-linked glycosylation include, e.g., the addition of N-acetyl-galactosamine (GalNAc), fucose, glucose, N-acetylglucosamine (GlcNAc), or mannose. GalNAc O-glycosylation is a particularly preferred PTM of the invention. GalNAc O-glycosylation is carried out by specific enzymes; for example, the addition of N-acetyl-galactosamine (GalNAc) to serine or threonine residues is carried out by the enzyme UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase (EC 2.4.1.41). Other types of O-glycosylation include αMannose, αFucose, βGlucose, βGlcNAc, and βXylose glycosylation. Also contemplated by the present invention are other forms of glycosylation, such as, e.g., N-linked glycosylation, hydroxyl-lysine glycosylation, and C-mannosylation. Biosynthesis and structures of these types of protein glycosylation are reviewed in Essentials of Glycobiology (2^nd edition, eds A. Varki, Cummings, Esko, Freeze, Stanley Bertozzi, Hart, Etzler. CSH Press 2009). It is to be understood that examples of PTM modification provided herein, such as O-glycosylation, are meant to be non-limiting examples which serve to prove the principal of the present methods (i.e., that the presently disclosed methods are useful for identifying PTM-modified peptides or peptide epitopes specifically recognized by disease-associated antibodies). The methods of the invention are also applicable, however, to proteins or peptides modified with other PTMs contemplated by the present invention, such as but not limited to phosphorylation, citrinylation, acetylation, and methylation.

[0068] In certain aspects of the invention, glycoproteins, glycopeptides and glycopeptide epitopes that are aberrantly glycosylated are provided. Examples of aberrant forms of O-glycans include, but are not limited to, truncated immature O-glycans such as Tn, T, and STn as well as non-sialylated and non-galactosylated core 2, 3 and 4 structures, shown in Table I below:

TABLE-US-00001 TABLE I Representative mucin-type O-glycan structures on normal and diseased cells¹ Type of Distribution Name Hapten Structure (+/-Ser/Thr or artificial linker) Glycoconjugate (main) No Tn GalNAcα1-O-Ser/Thr O-linked Cancer cells 1 STn NeuAcα2-3GalNAcα1-O-Ser/Thr O-linked Cancer cells 2 T Galβ1-3GalNAcα1-O-Ser/Thr O-linked Cancer cells 3 mSTa NeuAcA2-3Galβ1-3GalNAcα1-O-Ser/Thr O-linked Normal cells 4 mSTb NeuAcA2-3Galβ1-3[NeuAcα2-6]GalNAcα1-O- O-linked Normal cells 5 Ser/Thr Truncated C3 GlcNAcβ1-3GalNAcα1-O-Ser/Thr O-linked Cancer cells 6 Truncated C2 (NeuAc2-3).sub.+/-Galβ1-3[GlcNAcβ1-6]GalNAcα1-O- O-linked Cancer cells 7 Ser/Thr Truncated C4 GlcNAcβ1-3[GlcNAcβ1-6]GalNAcα1-O-Ser/Thr O-linked Cancer cells 8 Non-capped type1-C3 Galβ1-3GlcNAcβ1-3GalNAcα1-O-Ser/Thr O-linked Cancer cells 9 Non-capped type2-C2 (NeuAc2-3).sub.+/-Galβ1-3[Galβ1-4GlcNAcβ1- O-linked Cancer cells 10 6]GalNAcα1-O-Ser/Thr Non-capped type2-C4 (Galβ1-4)GlcNAcβ1-3[Galβ1-4GlcNAcβ1- O-linked Cancer cells 11 6]GalNAcα1-O-Ser/Thr GalNAcα-Tn GalNAcα1-3GalNAcα1-O-Ser/Thr O-linked Cancer cells 12 SA-type1-C3 NeuAcα2-3Galβ1-3GlcNAcβ1-3GalNAcα1-O- O-linked Cancer cells 13 Ser/Thr SLe^a-C3 NeuAcα2-3Galβ1-3[Fucα1-4]GlcNAcβ1- O-linked Cancer cells 14 3GalNAcα1-O-Ser/Thr LacDiNAc-C3 GalNAcβ1-3/4GlcNAcβ1-3GalNAcα1-O-Ser/Thr O-linked unknown 15 LacDiNAc-C2 Galβ1-3[GalNAcβ1-3/4GlcNAcβ1-6]GalNAcα1-O- O-linked unknown 16 Ser/Thr LacDiNAc-C4 GalNAcβ1-3/4GlcNAcβ1-3[GalNAcβ1- O-linked unknown 17 3/4GlcNAcβ1-6]GalNAcα1-O-Ser/Thr ¹Additional modifications such as capping terminal β3/4Gal residues with Fucα1-2 (blood group H) and Gal(NAc)α1-3 (blood group A,B) are generally produced by normal cells. Modifications such as 3/6-O--SO₃ of Gal and GalNAc, 9-O-acetylation of NeuAc, NeuGc instead of NeuAc, may also occur in normal and cancer cells.

[0069] In some aspects of the invention, the glycopeptide sequences of the invention may be derived from disease-associated proteins, such as, e.g., a glycoprotein specifically expressed on the cell surface of a cancer cell, but not on the surface of a non-malignant cell, or overexpressed on cancer cell surfaces, e.g., mucin 1 (MUC1) (GenBank Accession Nos. NP_002447 (SEQ ID NO: 1); NP_001018016 SEQ ID NO: 2); NP_001018017 (SEQ ID NO: 3); NP_001037855 (SEQ ID NO: 54); NP_001037856 (SEQ ID NO: 55); and NP_001037857 (SEQ ID NO: 56)); MUC16 (GenBank Accession No. NP_078966 (SEQ ID NO: 57); and mesothelin (MSLN) (GenBank Accession Nos. NP_037536 (SEQ ID NO: 58) and NP_005814 (SEQ ID NO: 59). In some embodiments, the glycoproteins are organ-specific glycoproteins. Examples of such glycoproteins include MUC16 (expressed in the female reproductive tract) and mesothelin, which is specifically expressed in the normal lung, but with cancer expression in ovarian and cervical cancers. Specific examples of other glycoproteins of the invention include, but are not limited to, human proteins predicted from the completed human genome containing one or more serine and/or threonine GalNAc O-glycosylation sites as predicted by the NetOGlyc algorithm.

[0070] Non-limiting examples of such proteins include cell membrane receptors such as low-density lipoprotein receptor precursor (LDLR) (GenBank Accesion No. NP_000518) (SEQ ID NO: 67)), very low-density lipoprotein receptor precursor (VLDLR) (GenBank Accession Nos. NP_001018066 (SEQ ID NO: 68) and NP_003374 (SEQ ID NO: 69)), and receptor tyrosine-protein kinase erbB-2 precursor (ERBB2) (GenBank Accession Nos. NP_004439 (SEQ ID NO: 70) and NP_001005862 (SEQ ID NO: 71)); chaperones such as heat shock 70 kDa protein 5 (HSPA5) (GenBank Accession No. NP_005338 (SEQ ID NO: 72) and heat shock 70 kDa protein 8 (HSPA8) (GenBank Accession No. NP_694881) (SEQ ID NO: 73) and NP_006588 (SEQ ID NO: 74); secreted cytokines, growth factors and glycoproteins such as interferon alpha-2 precursor (IFNA2) (GenBank Accession No. NP_000596) (SEQ ID NO: 75), interleukin-2 (IL2) (GenBank Accession No. NP_000577) (SEQ ID NO: 76), and inhibin beta B chain precursor (INHBB) (GenBank Accession No. NP_002184) (SEQ ID NO: 77); proteases such as matrix metalloproteinase-14 precursor (MMP14) (GenBank Accession No. NP_004986) (SEQ ID NO: 78); enzymes such as prostatic acid phosphatase precursor (ACPP) (GenBank Accession Noa. NP_001127666) (SEQ ID NO: 79) and NP_001090 (SEQ ID NO: 80); and mucins such as melanoma cell adhesion molecule (MCAM) (GenBank Accession No. NP_006491) (SEQ ID NO: 81).

[0071] In other embodiments, glycopeptide mutants are contemplated for use in the present invention. For example, a peptide library can be generated wherein each amino acid in the peptide sequence around one or more sites for PTM modification (e.g., O-glycosylation) is varied by any combination of the 20 amino acids used in human proteins to form additional peptide sequences (i.e. mutants) with the same PTM motif.

[0072] In certain embodiments, the present invention further relates to the design of glycopeptide libraries for screening disease-associated antibodies and/or for identifying disease-associated glycopeptides and/or glycopeptide epitopes. In certain aspects, a glycopeptide library of the invention is provided as a glycopeptide panel for identifying glycopeptides reactive with disease-associated auto-antibodies. A "glycopeptide panel" comprises at least two glycopeptides (or indeed entire glycoproteins) and may be used in an antibody binding assay of the invention. In certain aspects, methods for designing both the sequence of the glycopeptide and the specific site(s) for glycosylation are provided.

[0073] Preferably, glycopeptides of the glycopeptide libraries of the invention are from about 4 to about 50 amino acids in length. More preferably, such glycopeptides are from about 4 to about 25 amino acid residues in length.

[0074] In certain aspects, peptides are designed as fragments of a glycoprotein. A random peptide library encompassing peptides having between about 4 and about 50 amino acids, is provided, wherein each amino acid in each peptide is varied by all 20 amino acids used in human proteins to form different peptide sequences. These random peptides are PTM-modified, e.g., O-glycosylated, on at least one amino acid residue. In other aspects, the peptide library comprises peptides covering all serine and threonine residues in the predicted human protein sequences. In yet another aspect, peptides of the glycopeptide libraries comprise sequences derived from disease-associated proteins and/or organ-specific proteins, as described, supra.

[0075] In certain embodiments of the invention, a peptide suitable for testing is a potential acceptor of GalNAc O-glycosylation by polypeptide GalNAc-transferases if: 1) the peptide is a sequence derived from the human proteome, wherein the human proteome may be predicted from the human genome; and 2) the peptide comprises one or more serine and/or threonine residues. Preferably, such peptides are derived from proteins that have a signal sequence. It is also preferred that such peptides have sequences that, in vitro, serve as substrates for one or more human polypeptide GalNAc-transferases, such as, e.g., GalNAc-T1, T2, T3, or T4. The initiating enzymes for other types of O-glycosylation that attaches the first sugar residue to Serine and Threonine residues of polypeptides have been cloned and expressed recombinantly and hence these types of modifications are amiable to the same strategy for testing (Essentials of Glycobiology (2^nd edition, eds A. Varki, Cummings, Esko, Freeze, Stanley Bertozzi, Hart, Etzler. CSH Press 2009).

[0076] In certain aspects, it is preferred that the glycopeptides of the invention comprise sequences predicted by the NetOGlyc algorithm. The NetOGlyc algorithm has been developed to predict sites and proteins modified by O-glycosylation and this has been postulated to have a positive prediction rate of 76% (Julenius et al. 2005).

[0077] In other embodiments, peptides may be modified with other PTM modifications.

Glycopeptide Synthesis

[0078] In some aspects of the invention, the peptide sequence of the glycopeptides of the invention can be synthesized using standard chemical synthesis [Meldal M, Bock K. A general approach to the synthesis of O- and N-linked glycopeptides. Glycoconj J. 1994; 11(2):59-63]. For example, GalNAca-Ser/Thr-Fmoc, GlcNAcα-Ser/Thr-Fmoc, Manα-Ser/Thr, and Glcβ-Ser/Thr amino acids are commercially available (Sussex Inc (Canada)), and may be incorporated into the peptides during synthesis. The glycopeptides of the invention may be synthesized with and without an N-terminal linker for printing on epoxy or NHS-activated glass slides, respectively. In certain embodiments, glycopeptides may also be synthesized directly on microarray slides ("spot synthesis"). In other embodiments, peptides are previously synthesized and then immobilized or used in solution, according to a method of the present invention.

[0079] In certain aspects, the synthesized peptides or glycopeptides are O-glycosylated or further O-glycosylated using enzymatic synthesis of O-glycans at specific sites to form glycopeptides (Tarp and Clausen 2008).

[0080] In certain aspects of the present invention, the peptides may be glycosylated by a number of different methods, such as e.g., on-slide glycosylation, in solution glycosylation, or in vivo, e.g., by recombinant expression in appropriate host cells (Tarp et al. Glycobiology 2007).

[0081] Glycosylation may be achieved using one or more recombinant glycosyltransferases. such as, e.g., recombinant polypeptide GalNAc-transferases (e.g. GalNAc-T2, -T3 and -T4). See, e.g., U.S. Pat. No. 5,876,716 by Hansen and U.S. Pat. No. 6,465,220 by Hassan; see also, Bennett et al. 1998; Bennett et al. 1996; and White et al. 1995.

[0082] Further expansion may also be achieved using a recombinant sialyltransferase, ST6GalNAc-II, to produce STn glycoforms (structure no. 2, Table I) of the GalNAc glycopeptides. T glycoforms may be produced by a recombinant Drosophila core1 β3galactosyltransferase, truncated core3 glycoforms (structure no. 6, Table I) may be produced using a recombinant human β3GlcNAc-transferase (Iwai et al. 2002), and non-capped type1-core3 glycoforms (structure no. 9, Table I) may be produced using β3Gal-T5 (see, U.S. Pat. No. 7,332,279).

[0083] In other aspects, the glycopeptides and/or glycoproteins of the invention may be derived from recombinant or isolated glycopeptides or glycoproteins and further glycosylated or modified according to the methods of the present invention. For example, commercially available glycoprotein arrays (available, e.g., from Invitrogen), may be treated with exoglycosidases, e.g., neuraminidase, βgalactosidase, βN-acetylglucosaminidase and other enzymes, in order to expose cancer-associated glycans, e.g., Tn or T, to form glycopeptides according to the present invention.

[0084] In certain aspects of the invention, the O-glycan is preferably positioned in the center of the peptide. While not intending to be bound by a specific theory, positioning the O-glycan in the center of the peptide may facilitate proper presentation of the O-glycan for specific antibody binding to a glycopeptides epitope. In certain aspects, for example, in a glycopeptide of 20 amino acid residues in length, the O-glycan is preferably attached to a serine or threonine amino acid residue placed at a site from about residue 6 to about residue 15 and more preferably from about residue 8 to about residue 13. For example, in the present Examples, glycopeptides were designed with a single GalNAc at position 12 (of the 20-mer) to allow optimal exposure of peptide sequence flanking the O-glycan, taking into consideration that the glycopeptides will be covalently linked primarily through the N-terminal amino acid.

[0085] In certain aspects, the O-glycans to be presented on a glycopeptide library of the invention may include all known O-glycan structures. In preferred embodiments, the glycopeptides comprise glycans known to be disease-associated, such as Tn, STn, T, Truncated C3, Truncated C2, Truncated C4, non-capped type1-C3, non-capped type2-C2, non-capped type2-C4, GalNAca-Tn, SA-type1-C3, SLea-C3, LacDiNAc-C3, LacDiNAc-C2, and LacDiNAc-C4 (see Table I for additional non-limiting examples of such glycans). Further the O-glycans may be substituted with sulfation or other immunogenic substitutions including acetylation and artificial chemical groups.

Screening Assays

[0086] In certain embodiments, the present invention provides methods for identifying glycopeptides and/or glycopeptide epitopes reactive with disease-associated auto-antibodies using an assay comprising one or more glycopeptide panels. Preferably, such auto-antibodies bind glycopeptide epitopes through recognition of both the peptide portion and the glycan portion, but not through recognition of either the peptide or the glycan alone. The panel is contacted with a sample containing the disease-associated antibodies (e.g., sera obtained from an individual with the disease).

[0087] In a specific embodiment, the invention provides a method for identifying glycopeptides reactive with cancer-associated auto-antibodies, wherein the method comprises: (a) providing a panel comprising glycopeptides having a peptide portion and a glycan portion; (b) contacting the panel with an antibody-containing a sample from a patient with cancer; and (c) identifying glycopeptides in the panel that (i) are selectively recognized by antibodies in the sample, but not by antibodies in a control sample, and (ii) are recognized by such antibodies in the sample through recognition of both the peptide portion and the glycan portion, but not through recognition of either the peptide or the glycan alone.

[0088] Non-limiting examples of suitable antibody-containing samples for the assays of the present invention include serum, plasma, body fluids such as milk, saliva, mucosal secretions, feces, urine, cells and tissues, and any antibody preparations thereof.

[0089] Further, in order to develop the glycopeptide screening assays of the present invention, a control sample is used. By "control sample", it is meant a sample containing pooled sera obtained from apparently disease-free or healthy individuals (i.e., individuals who do not have the disease-associated antibodies in their serum because they do not have the relevant disease), or, it is meant multiple control samples, wherein each sample is obtained from a single apparently healthy (disease-free) individual, and then data obtained in the assay for the tested control samples are compared in order to exclude any samples from control individuals who are suspected to in fact not be healthy, based on the presence of auto-antibodies not present in a statistically significant fraction of the control population.

[0090] In certain aspects, the present invention provides methods for identifying cancer-associated glycopeptide epitopes, wherein the epitope includes both part of the glycan of the glycopeptide and part of the peptide amino acid sequence. Such methods can comprise the following steps: (a) providing a panel comprising glycopeptides having a peptide portion and a glycan portion; wherein the peptide amino acid sequences of the glycopeptides are serially shifted 1-5 residues in either direction, to provide a series of overlapping peptide sequences; (b) contacting the panel with an antibody-containing sample from a patient with cancer; (c) identifying glycopeptides in the panel that (i) are selectively recognized by antibodies in the sample, but not by antibodies in a control sample, and (ii) are recognized by such antibodies in the sample through recognition of both the peptide portion and the glycan portion, but not through recognition of either the peptide or the glycan alone; and (d) mapping the minimal glycopeptide epitope based on the pattern of antibody binding to the overlapping glycopeptide sequences.

[0091] "Epitope mapping" may be carried out as follows: A glycopeptide epitope comprising e.g., a 20-mer glycopeptide with a single glycan attached to position 12 can be "mapped" in terms of peptide sequence requirement by synthesis of a panel of 20-mer glycopeptides in which each amino acid around the glycan site is modified one by one to an amino acid different from the one present in the identified glycopeptide, e.g. alanine or valine. Analysis of antibody binding to this panel of "walking" alanine or valine residues through the peptide sequence will demonstrate which residues abrogate binding, and thus provide information of the necessary peptide sequence backbone around the glycosylation site required for antibody binding.

[0092] In certain aspects of the invention, the minimum glycopeptide epitope is preferably about 2 to about 7 amino acid residues in length, but can extend to up to about 15 residues, and comprises a portion of an O-glycan modifying the glycopeptide epitope. In certain embodiments, the minimal glycopeptide epitope spans about 3 to about 4, about 2 to about 3, or about 1 to about 2 amino acid residues on each side of the O-glycan that is part of the minimal epitope identified by the methods of the present invention. In other embodiments, a greater number of amino acids may be present on one side of the O-glycan compared to the number of amino acids of the minimal epitope on the other side of the O-glycan. The minimum glycopeptide epitope can be, for example, a "minimum cancer-associated glycopeptide epitope," which is the minimum glycopeptide epitope identified by the methods of the present invention to be specifically recognized by a cancer-associated auto-antibody.

[0093] In certain embodiments, the epitope structure of the glycopeptide epitopes identified by the present methods is determined by mass spectrometry.

[0094] In some aspects, determining whether glycopeptides and/or glycopeptide epitopes are selectively recognized by antibodies in a sample through recognition of both the peptide portion and the glycan portion, wherein the antibodies do not recognize either portion alone, may be achieved as follows: an additional panel may be provided, in which the unglycosylated forms of the glycopeptides are provided (i.e., the corresponding peptides) and antibody binding is determined. In order for a glycopeptide to be selectively recognized by an antibody in the sample, antibody binding to the corresponding peptide should be partially or preferably completely abrogated. Preferably, antibody binding to the corresponding peptide is diminished by at least about 40%, preferably at least about 50%, more preferably at least about 60%, even more preferably at least about 70%, still more preferably at least about 80%, more preferably at least about 85%, still more preferably at least about 90%, and most preferably 95% to 100% compared to antibody binding to the glycopeptide.

[0095] In other aspects of the present invention, methods of determining the presence of auto-antibodies binding O-glycopeptides, may also comprise the step of contacting the panel with a peptide inhibitor in order to exclude antibodies that recognize the non-glycosylated form of the glycopeptide. Peptide inhibitors are typically a peptide of the same amino acid sequence as the glycopeptide, however, without any glycosylations.

[0096] In other embodiments, antibodies that recognize only the hapten glycan or the glycan-conjugated to a different peptide or artificial carrier may be excluded, in order to prevent identification of epitopes that consist only of the glycan hapten. This can be achieved in several ways. For example, in certain embodiments, hapten-specific antibodies may be removed from the sample prior to analysis.

[0097] In some embodiments, hapten-specific antibodies may be removed by affinity chromatography with an appropriate resin with covalently linked carbohydrate haptens, or by inhibition with carbohydrate haptens in solution during binding assays. An example of preferable resin is GalNAc-Sepharose or other appropriate resins that would be able to bind anti-Tn hapten antibodies. Other preferable resins are GlcNAc-Sepharose, Man-Sepharose, Glc-Sepharose or Fuc-Sepharose and without limitations corresponding di- and trisaccharides in the biosynthetic pathways bound resins.

[0098] In other embodiments, hapten-specific antibodies may excluded from binding to glycopeptides in the binding assays of the present invention using a carbohydrate inhibitor. In some embodiments, the carbohydrate inhibitor comprises a normal occurring O-glycan such as sialylated or fucosylated core 2,3, or 4 O-glycan, as is typically present in normal cells. In some aspects, the carbohydrate inhibitor may be Tn, STn, T or other truncated O-glycan structures based on core 3 or 4. Also preferred are polyvalent PAA (polyacrylamide) conjugates (GlycoTech, US) of the aforementioned carbohydrates. Still in another embodiment, the carbohydrate inhibitor is a monosaccharide such as GalNAc, GlcNAc, Gal, Glc, Fuc, Man, Xyl and NeuAc. It will be apparent to the skilled artisan that other combinations of carbohydrates will have the same effect.

[0099] The above-described methods, as well as other methods that may be readily determined by a skilled artisan and used to achieve the same effect, may be used to exclude antibodies in a sample that do not specifically recognize the glycopeptide (i.e., through both the peptide and the glycan). In certain embodiments of the invention, exclusion of such antibodies from the analysis is necessary in order to identify the glycopeptides and glycopeptide epitopes that are specifically recognized by disease-associated antibodies.

[0100] In certain embodiments, a peptide inhibitor and/or an O-glycan inhibitor may be immobilized on a solid support, which is used to remove antibodies that interact with the peptide inhibitor and/or with the O-glycan carbohydrate inhibitor from a sample, prior to use of the sample according to a method of the present invention.

[0101] In some aspects, the screening assays of the present invention involve immobilized glycopeptides; however, in solution assays such as polarization, inhibition and competitive binding assays are also contemplated for use in the present invention. [See, e.g., Smith D S, Eremin S A. Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules. Anal Bioanal Chem. 2008; 391(5):1499-507.]

[0102] Non-limiting examples of screening assays involving immobilized glycopeptides contemplated for use in the present invention include antibody-binding assays, such as enzyme-linked immunosorbent assay (ELISA), multiplex bead arrays (see, Elshal et al., (2006) Methods; 38(4):317-323); BiaCore SPR analysis where binding affinities can be evaluated and microarray platforms.

[0103] In a preferred embodiment, glycopeptide libraries of the invention are immobilized on microarray slides. The glycopeptides may be printed on microarray slides, such as, e.g., Corning, Scineion or Nexterion® Slide H or Schott Nexterion® Slide H MPX 16 (Schott A G, Mainz, Germany) by JPT (Germany). Printing may be carried out using MicroGrid, ArraylT or similar according to the methods described in (Blixt O, Head S, Mondala T, Scanlan C, Huflejt M E, Alvarez R, Bryan M C, Fazio F, Calarese D, Stevens J, Razi N, Stevens D J, Skehel J J, van Die I, Burton D R, Wilson I A, Cummings R, Bovin N, Wong C H, Paulson J C. Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc Natl Acad Sci USA. 2004; 101(49):17033-8)

[0104] Microarray slides of the invention may also be prepared by companies such as Schott (Louisville, Ky.), Arraylt Corp. (Sunnyvale, Calif.), or Scineon (Germany).

[0105] In certain embodiments, assays for large scale screening include, e.g., multiplex bead or array formats where many targets can be assayed simultaneously with very little consumption of antibody. For example, a small volume (50 μl) of an antibody-containing sample (e.g. serum or diluted serum) may be incubated with a solution of beads, wherein each bead is coated with a specific glycopeptide, and each bead possesses a distinguishing characteristic (e.g., size) that allows it to be differentiated from other beads in the sample. If an antibody is present in the sample that recognizes a glycopeptide on one of the beads, it will bind to the bead. Then, specific binding of antibodies to each glycopeptide in the sample may be detected using a detecting reagent, such as, e.g., biotinylated anti-Ig antibody followed by fluorescently-labeled streptavidin. The concentration of antibody in the sample that is specific for each glycopeptide on the bead may then be quantified using a bead analyzer, such as, e.g., the Luminex® 200® System (Invitrogen).

[0106] In yet another embodiment of the invention, the detection of auto-antibodies can be limited to distinct human Ig isotypes and subclasses. Most human natural carbohydrate antibodies are of IgM isotype and it is the detection of auto-antibodies of IgG isotype and subclasses is preferred. More specifically, detection of binding of human IgG antibodies, or IgG1, 2, 3, and 4 individually, with appropriate anti-human antibodies avoid most reactivity with truncated O-glycan haptens such as Tn, STn, T core3, and other truncated structures to which IgM antibodies are found in control samples.

Methods of Diagnosing

[0107] In some aspects, the present invention provides methods for diagnosing a patient with a disease. For example, disease-associated glycopeptides and glycopeptide epitopes identified by the methods of the present invention may be used for detection of disease-associated auto-antibodies with the purpose of determining diagnosis and/or prognosis.

[0108] For example, in certain embodiments, a patient may be diagnosed as having cancer or as not having cancer, based on the presence or absence, respectively, of specific, cancer-associated glycopeptide-reactive auto-antibodies. In a specific embodiment, the method comprises contacting an antibody-containing sample from a patient with a panel comprising peptides, wherein at least a plurality of the peptides are glycopeptides. Further, each glycopeptide comprises a glycopeptide epitope that has been previously determined (i) to be selectively recognized by a subset of antibodies in sera from cancer patients, which subset recognizes neither (a) the corresponding naked peptides of said panel when not glycosylated; nor (b) the corresponding glycan when not bound to said peptide; and (ii) not to be recognized by antibodies in control sera. It is then determined if antibodies in the sample are bound to glycopeptides of said panel; and concluded either that the patient has cancer if the sample comprises antibodies that bind to at least one of the glycopeptides in the panel; or that the patient does not have cancer if the sample does not comprise antibodies that bind to at least one glycopeptide in the panel.

[0109] In certain aspects, the specific type of cancer that may be diagnosed or treated by a method of the present invention without limitation may be selected from the group consisting of breast cancer, colon cancer, ovarian cancer, cervical cancer, pancreatic cancer, prostatic cancer, liver cancer, kidney cancer, brain cancer, hematological cancers, testis cancer, head and neck cancers, and lung cancer.

[0110] In yet another embodiment, the diagnostic panel comprises glycopeptides comprising one or more and preferably at least 8 of the amino acid sequences selected from the group consisting of SEQ ID NOs. 15, 36, 49, and 82-146.

[0111] In yet another specific embodiment, the diagnostic panel comprises glycopeptides comprising one or more of the amino acid sequences selected from the group consisting of SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146). Furthermore, these glycopeptides may be O-glycosylated at one or more sites according to the methods of the present invention.

[0112] In yet another embodiment, the diagnostic panel comprises at least 8 glycopeptides, for example between about 8 and about 30 glycopeptides, such as 10, 12, 15, or 20 glycopeptides. A number of glycopeptides in excess of 30 is also within the invention. The upper limit of glycopeptides in a panel is limited by practical considerations (e.g., how many glycopeptides can fit on a substrate) or cost-benefit considerations. Preferably the glycopeptides are selected from the group consisting of SEQ ID NOs 15, 36, 49, and 82-146.

[0113] In yet other aspects, a patient may be diagnosed as having an autoimmune disease. In certain embodiments, the autoimmune disease is selected from the group consisting of coeliac disease, type I diabetes, multiple sclerosis, thyroiditis, Grave's disease, systemic lupus erythematosus, scleroderma, psoriasis, rheumatoid arthritis, alopecia greata, ankylosing spondylitis, Churg-Strauss Syndrome, autoimmune hemolytic anemia, autoimmune hepatitis, Behcet's disease, Crohn's disease, dermatomyositis, glomerulonephritis, Guillain-Barre syndrome, inflammatory bowel disease (IBD), lupus nephritis, myasthenia gravis, myocarditis, pemphigus/pemphigoid, pernicious anemia, polyarteritis nodosa, polymyositis, primary biliary cirrhosis, rheumatic fever, sarcoidosis, Sjogren's syndrome, ulcerative colitis, uveitis, vitiligo, and Wegener's granulomatosis.

[0114] Thus, in some embodiments, a diagnostic panel of the invention comprises glycopeptide epitopes determined by the methods of the present invention to be associated with an autoimmune disease. For example, in certain embodiments, a patient may be diagnosed as having an autoimmune disease or as not having an autoimmune disease, based on the presence or absence, respectively, of specific, autoimmune disease-associated glycopeptide-reactive auto-antibodies. In a specific embodiment, the method comprises contacting an antibody-containing sample from a patient with a panel comprising peptides, wherein at least a plurality of the peptides are glycopeptides. Further, each glycopeptide comprises a glycopeptide epitope that has been previously determined (i) to be selectively recognized by a subset of antibodies in sera from patients with a specific autoimmune disease, which subset recognizes neither (a) the corresponding naked peptides of said panel when not glycosylated; nor (b) the corresponding glycan when not bound to said peptide; and (ii) not to be recognized by antibodies in control sera. It is then determined if antibodies in the sample are bound to glycopeptides of said panel; and concluded either that the patient has the autoimmune disease if the sample comprises antibodies that bind to at least one of the glycopeptides in the panel; or that the patient does not have the autoimmune disease if the sample does not comprise antibodies that bind to at least one glycopeptide in the panel.

Preparation of Antibodies

[0115] Other aspects of the present invention are antibodies prepared using one or more glycopeptides identified using the methods of the present invention, methods for preparation of these antibodies, and the use of such antibodies in therapy and diagnosis.

[0116] Yet another aspect of the present invention is a method for the preparation of hybridoma cells, which secrete monoclonal antibodies specific for the glycopeptides of the invention. One such method involves immunizing a suitable mammal with a glycopeptide of the invention; fusing antibody-producing cells of the mammal with cells of a continuous cell line; the hybrid cells obtained in the fusion are cloned; and cell clones secreting the desired antibodies are selected.

[0117] Still another aspect is a monoclonal antibody selected from the group consisting of: a monoclonal antibody produced by the hybridoma cells prepared by the method described above; and a monoclonal antibody prepared by molecular display techniques, such as mRNA display, ribosome display, phage display and covalent display against a glycopeptide of the invention.

[0118] For preparation of monoclonal antibodies, any technique that provides for the production of antibody molecules by continuous cell lines in culture may be used. These include but are not limited to the hybridoma technique originally developed by Kohler and Milstein (Nature, 1975; 256:495-497), as well as the trioma technique, the human B-cell hybridoma technique (Kozbor et al., Immunology Today, 1983; 4:72, Cote et al., Proc. Natl. Acad. Sci. U.S.A., 1983; 80:2026-2030), and the EBV-hybridoma technique to produce human monoclonal antibodies (Cole et al., in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96, 1985). In an additional embodiment of the invention, monoclonal antibodies can be produced in germ-free animals (International Patent Publication No. WO 89/12690, published 28 Dec. 1989).

[0119] According to the invention, techniques described for the production of single chain antibodies (U.S. Pat. Nos. 5,476,786 and 5,132,405 to Huston; U.S. Pat. No. 4,946,778) can be adapted to produce glycopeptide-specific single chain antibodies. Indeed, these genes can be delivered for expression in vivo to, e.g., express a glycopeptide-specific antibody. An additional embodiment of the invention utilizes the techniques described for the construction of Fab expression libraries (Huse et al., Science, 1989; 246:1275-1281) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity for a glycopeptide or glycopeptide epitope of the invention.

[0120] Antibody fragments which contain the idiotype of the antibody molecule can be generated by known techniques. For example, such fragments include but are not limited to: the F(ab')2 fragment which can be produced by pepsin digestion of the antibody molecule; the Fab' fragments which can be generated by reducing the disulfide bridges of the F(ab')2 fragment, and the Fab fragments which can be generated by treating the antibody molecule with papain and a reducing agent.

[0121] In the production of antibodies, screening for the desired antibody can be accomplished by techniques known in the art, e.g., radioimmunoassay, ELISA (enzyme-linked immunosorbant assay), "sandwich" immunoassays, immunoradiometric assays, gel diffusion precipitin reactions, immunodiffusion assays, in situ immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), Western blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.

[0122] In other embodiments, alternative techniques such as mRNA display, ribosome display, phage display and covalent display may also be used. These are all display techniques where a peptide library is selected against the glycopeptide. Such techniques can e.g. be used to identify humanized or fully human antibodies.

[0123] In a preferred embodiment, the monoclonal antibody binds an O-glycoprotein containing the O-glycopeptide epitope on cancer cells but not on a non-malignant counterpart.

[0124] In another preferred embodiment, the monoclonal antibody binds preferentially to the O-glycoprotein that is aberrantly glycosylated and expressed on cancer cells.

[0125] In still another embodiment, the monoclonal antibody binds to or at least interacts directly with the O-glycopeptide.

[0126] In a preferred embodiment, the antibody prepared using a glycopeptide of the invention is humanized or fully human, such as to decrease the immunogenicity of the antibody in humans. This is typically desirable if the antibody is used as a therapeutic. However, in some situations a rapid clearance may be desired, wherefore non-humanized antibodies are also of interest as therapeutics. One such situation can, e.g., be when administering antibody conjugates where antibodies are coupled to toxins or radioisotopes. Such conjugated antibodies should either find their target rapidly or be cleared as they have a general toxic effect. Thus, one embodiment of the invention is conjugated antibodies. Non-limiting examples of antibody conjugates include radioisotopes, such as ¹³¹I, ⁹⁰Y, ¹⁷⁷Leutitium (¹⁷⁷Lu) and ⁶⁷Copper (⁶⁷Cu); toxins, such as the fungal toxin maytansanoid (DM-1); and antibiotics, such as e.g., calicheamicin [See, Ross et al. Antibody-based therapeutics: Focus on prostate cancer. Cancer and Metastasis Reviews 24: 521-537, 2005].

Compositions and Uses

[0127] In certain embodiments, the present invention provides compositions comprising one or more disease-associated glycopeptides and/or glycopeptide epitopes identified by the methods of the present invention. In certain aspects, such disease-associated glycopeptides comprise epitopes specifically recognized and bound by disease-associated antibodies.

[0128] In a specific embodiment, a composition of the invention comprises one or more of a glycoprotein or glycopeptide comprising an amino acid sequence selected from the group consisting of SEQ IDs 15, 36, 49, and 82-146. Preferably, the composition comprises one or more of a glycoprotein or glycopeptides comprising an amino acid sequence selected from the group consisting of SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146). In certain embodiments, such glycoproteins or glycopeptides are further modified at at least one amino acid residue, with an O-glycan. Preferably, the amino acid residue is serine or threonine residue.

[0129] In some aspects, the invention provides methods for eliciting an immune response in an individual, wherein the immune responses is specific for one or more glycopeptides or glycoproteins identified by the methods of the present invention.

[0130] One aspect of the invention is a method of treating cancer or an autoimmune disease comprising administering a pharmaceutical composition of the invention. In a specific embodiment, a cancer patient is treated by eliciting an anti-cancer immune response that attacks cancer cells (e.g., tumors). In any of the aspects of the invention, the immune response elicited by a pharmaceutical composition of the invention may be an adaptive T and/or B cell response (e.g., either a cytotoxic T cell response or an antibody response, or both) directed against the cancer cells, which results in reduction or elimination of the cancer cells.

[0131] "Treating" or "treatment" of a state, disorder or condition includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human or other mammal that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition, (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof, or (3) relieving the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms.

[0132] The benefit to an individual to be treated is either statistically significant or at least perceptible to the patient or to the physician.

Pharmaceutical Compositions and Administration

[0133] While it is possible to use a composition provided by the present invention for therapy as is, it may be preferable to administer it in a pharmaceutical formulation, e.g., in admixture with a suitable pharmaceutical excipient, diluent, or carrier selected with regard to the intended route of administration and standard pharmaceutical practice. Accordingly, in one aspect, the present invention provides a pharmaceutical composition or formulation comprising at least one composition of the invention, or a pharmaceutically acceptable derivative thereof, in association with a pharmaceutically acceptable excipient, diluent, and/or carrier. The excipient, diluent and/or carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0134] The compositions of the invention can be formulated for administration in any convenient way for use in human or veterinary medicine.

Pharmaceutical Carrier

[0135] The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water or aqueous solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions. Alternatively, the carrier can be a solid dosage form carrier, including but not limited to one or more of a binder (for compressed pills), a glidant, an encapsulating agent, a flavorant, and a colorant. Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin (1990, Mack Publishing Co., Easton, Pa. 18042).

Vaccines

[0136] In certain aspects, a disease-associated glycopeptide or glycoprotein of the present invention is provided in a vaccine. Thus, in certain aspects, such a disease-associated glycopeptide or glycoprotein is provided as an "immunogen" for inducing an immune response. In some aspects, a vaccine of the present invention is useful for treating cancer by inducing an "anti-cancer immune response." Preferably, a vaccine of the present invention in effective for inducing an adaptive immune response that selectively target cancer cells and has minimal reactivity with normal cells.

[0137] In certain embodiments, the glycopeptide epitopes identified by the methods of the present invention are targets for spontaneously-induced human auto-antibodies. Thus, it is evident that these epitopes are not generally covered by immunological tolerance. Accordingly, in certain aspects of the invention, a vaccine comprising a glycopeptide immunogen identified by the present methods does not require an adjuvant.

[0138] In other embodiments, a vaccine comprising a glycopeptide immunogen of the invention may additionally contain adjuvants to induce or enhance the desired immune response, such as, e.g., an anti-cancer immune response. Exemplary adjuvants include, but are not limited to, cholera toxin, fragments and mutants or derivatives with adjuvant properties, E. coli heat-labile enterotoxin, fragments and mutants or derivatives with adjuvant properties, oil-in-water and water-in-oil emulsions, toll-like receptor ligands such as muramyl dipeptide, E. coli LPS, oligonucleotides comprised of unmethylated DNA, poly I:C, lipoteichoic acid, peptidoglycan. Enterotoxins and their adjuvant active derivatives such as cholera toxin, heat-labile E. coli enterotoxin, pertussis toxin, shiga toxin and analogs. Other adjuvants can be used such as complete Freund's adjuvant, incomplete Freund's adjuvant, saponin, mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil or hydrocarbon emulsions, keyhole limpet hemocyanins, and potentially useful human adjuvants such as N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-nor-muramyl-L-alanyl-D-isoglutamine, N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(1'-2'-dipalmitoyl-s- n-glycero-3-hydroxyphosphoryloxy)-ethylamine, BCG (bacille Calmette-Guerin) and Corynebacterium parvum. An adjuvant can serve as a tissue depot that slowly releases the antigen and also as a lymphoid system activator that non-specifically enhances the immune response (Hood et al., Immunology, Second Ed., 1984, Benjamin/Cummings: Menlo Park, Calif., p. 384). Where the vaccine is intended for use in human subjects, the adjuvant should be pharmaceutically acceptable.

Formulations

[0139] The compositions, vaccines and formulations of the present invention may comprise pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions include diluents of various buffer content (e.g., Tris-HCl, acetate, phosphate), pH and ionic strength; additives such as detergents and solubilizing agents (e.g., Tween 80, Polysorbate 80), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite), preservatives (e.g., Thimersol, benzyl alcohol) and bulking substances (e.g., lactose, mannitol); incorporation of the material into particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, etc. or into liposomes. Hylauronic acid may also be used. See, e.g., Remington's Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, Pa. 18042) pages 1435 1712 which are herein incorporated by reference.

[0140] Preparations according to this invention for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, or emulsions. Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. Such dosage forms may also contain adjuvants, preserving, wetting, emulsifying, and dispersing agents. The pharmaceutical compositions may be sterilized by, for example, filtration through a bacteria retaining filter, by incorporating sterilizing agents into the compositions, by irradiating the compositions, or by heating the compositions. They can also be manufactured using sterile water, or some other sterile injectable medium, immediately before use.

Administration and Dosage

[0141] The compositions (e.g., pharmaceutical or vaccine compositions) and formulations of the present invention can be administered topically, parenterally, orally, by inhalation, as a suppository, or by other methods known in the art. The term "parenteral" includes injection (for example, intravenous, intraperitoneal, epidural, intrathecal, intramuscular, intraluminal, intratracheal or subcutaneous). The preferred routes of administration are subcutaneous and intravenous.

[0142] The compositions and formulations of the present invention may be administered to an animal, preferably a mammal, and most preferably a human.

[0143] The dosage of the compositions or formulations of the present invention will vary widely, depending upon the nature of the disease, the patient's medical history, age, body weight, sex, sensitivity, the frequency of administration, the manner and route of administration, the clearance of the agent from the host, dosage period, drugs used in combination, and the like. The initial dose may be larger, followed by smaller maintenance doses.

[0144] For any composition or formulation used in the methods of the invention, the therapeutically effective dose can be estimated initially from animal models. Dose-response curves derived from animal systems are then used to determine testing doses for the initial clinical studies in humans. In safety determinations for each composition, the dose and frequency of administration should meet or exceed those anticipated for use in the clinical studies.

[0145] Toxicity and therapeutic efficacy of the compositions, vaccines, and formulations of the invention can be determined by standard pharmaceutical procedures in experimental animals, e.g., by determining the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (the dose therapeutically effective in 50% of the population). The dose ratio between therapeutic and toxic effects is the therapeutic index and it can be expressed as the ratio ED₅₀/LD₅₀. Compositions that exhibit large therapeutic indices are preferred.

[0146] The data obtained from the animal studies can be used in formulating a range of doses for use in humans. The therapeutically effective doses of in humans lay preferably within a range of circulating concentrations that include the ED₅₀ with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. Ideally, a single dose of each drug should be used daily.

[0147] The compositions of the invention will typically contain an effective amount of the compositions for achieving the desired effect. As used herein, an "effective amount of a glycopeptide" is an amount that elicits the desired response upon administration, e.g., an amount that elicits an immune response in a mammal.

[0148] Administration of the compositions or formulations of the invention may be once a day, twice a day, or more often, but frequency may be decreased during a maintenance phase of the disease or disorder, e.g., once every second or third day instead of every day or twice a day. The dose and the administration frequency will depend on the clinical signs, which confirm maintenance of the remission phase, with the reduction or absence of at least one or more preferably more than one clinical signs of the acute phase known to the person skilled in the art. More generally, dose and frequency will depend in part on recession of pathological signs and clinical and subclinical symptoms of a disease condition or disorder contemplated for treatment with the present compounds.

[0149] The appropriate dose and dosage times under certain conditions can be determined by the test based on the above-described indices but may be refined and ultimately decided according to the judgment of the practitioner and each patient's circumstances (age, general condition, severity of symptoms, sex, etc.) according to standard clinical techniques.

[0150] In certain embodiments of the invention, use of an appropriate vaccine design and immunization scheme will therefore elicit immunity to a glycopeptide epitope of the immunogen and efficiency of the immunization can be monitored by immunoassays, e.g., by detecting the presence of immunogen-specific antibodies in an antibody-containing sample from an immunized patient by ELISA or other assay suitable for detecting antigen-specific antibodies.

[0151] Keeping the above description in mind, typical dosages of the glycopeptide-containing compositions of the invention are 5-50 μg glycopeptides conjugated to e.g. KLH (keyhole-Limpet Hemocyanin) given subcutaneously 3-5 times with 2-3 weeks apart. Maintenance vaccine could be extended with monthly or bi-monthly dosing for extended periods.

Kits

[0152] In some embodiments, the invention relates to a kit comprising one or more glycopeptides identified by the present methods. In certain aspects the kit comprises a panel of two or more glycopeptides identified by the present methods. In other aspects, the kit further provides instructions for use. In a specific embodiment, the kit provides a diagnostic assay for diagnosing cancer, comprising a panel of cancer-associated glycopeptides, assay buffers, and instructions for use.

[0153] In accordance with the present invention, there may be employed conventional molecular biology, microbiology, recombinant DNA, immunology, cell biology and other related techniques within the skill of the art. See, e.g., Sambrook et al., (2001) Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, N.Y.; Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, N.Y.; Ausubel et al., eds. (2005) Current Protocols in Molecular Biology. John Wiley and Sons, Inc.: Hoboken, N.J.; Bonifacino et al., eds. (2005) Current Protocols in Cell Biology. John Wiley and Sons, Inc.: Hoboken, N.J.; Coligan et al., eds. (2005) Current Protocols in Immunology, John Wiley and Sons, Inc.: Hoboken, N.J.; Coico et al., eds. (2005) Current Protocols in Microbiology, John Wiley and Sons, Inc.: Hoboken, N.J.; Coligan et al., eds. (2005) Current Protocols in Protein Science, John Wiley and Sons, Inc.: Hoboken, N.J.; Enna et al., eds. (2005) Current Protocols in Pharmacology John Wiley and Sons, Inc.: Hoboken, N.J.; Hames et al., eds. (1999) Protein Expression: A Practical Approach. Oxford University Press: Oxford; Freshney (2000) Culture of Animal Cells: A Manual of Basic Technique. 4th ed. Wiley-Liss; among others. The Current Protocols listed above are updated several times every year.

EXAMPLES

[0154] The present invention is described further below in working examples which are intended to further describe the invention without limiting the scope therein.

Example 1

Development of a Cancer-Associated O-Glycopeptide Library for Microarray Display

[0155] GalNAc-Ser/Thr-Fmoc amino acids were synthesized by Sussex Inc (Canada) and 20-mer peptides and corresponding glycopeptides with one GalNAc O-glycan synthesized and printed on epoxy activated glass slides by JPT Peptide Technologies (Berlin, Germany). A total of 48 paired peptides and 48 GalNAc-glycopeptides were synthesized and printed (Table II).

TABLE-US-00002 TABLE II Synthetic 96 paired glycopeptide 20-mer library derived from confirmed O-glycoproteins* SEQ ID Peptide SEQ ID Peptide NO: No. NO: No. 4 1 TSAPDTRPAPGSTAPPAHGV 29 49 CDKDKTSTVAPTIHTTVPSP 4 2 TSAPDTRPAPGSTAPPAHGV 30 50 DKTSTVAPTIHTTVPSPTTT 5 3 SAPDTRPAPGSTAPPAHGVT 30 51 DKTSTVAPTIHTTVPSPTTT 5 4 SAPDTRPAPGSTAPPAHGVT 31 52 KTSTVAPTIHTTVPSPTTTP 6 5 PGSTAPPAHGVTSAPDTRPA 31 53 KTSTVAPTIHTTVPSPTTTP 6 6 PGSTAPPAHGVTSAPDTRPA 32 54 TVAPTIHTTVPSPTTTPTPK 7 7 GSTAPPAHGVTSAPDTRPAP 32 55 TVAPTIHTTVPSPTTTPTPK 7 8 GSTAPPAHGVTSAPDTRPAP 33 56 APTIHTTVPSPTTTPTPKEK 8 9 PPAHGVTSAPDTRPAPGSTA 33 57 APTIHTTVPSPTTTPTPKEK 8 10 PPAHGVTSAPDTRPAPGSTA 34 58 PTIHTTVPSPTTTPTPKEKP 9 11 FYLAMPFATPMEAELARRSL 34 59 PTIHTTVPSPTTTPTPKEKP 10 12 KFSEFWDLDPEVRPTSAVAA 35 60 TIHTTVPSPTTTPTPKEKPE 11 13 PLVEQGRVRAATVGSLAGQP 35 61 TIHTTVPSPTTTPTPKEKPE 11 14 PLVEQGRVRAATVGSLAGQP 36 62 TPTPKEKPEAGTYSVNNGND 12 15 LWSLCWSLAIATPLPPTSAH 36 63 TPTPKEKPEAGTYSVNNGND 12 16 LWSLCWSLAIATPLPPTSAH 37 64 ACLAVSAGPVPTPPDNIQVQ 13 17 LEACVIQGVGVTETPLMKED 37 65 ACLAVSAGPVPTPPDNIQVQ 13 18 LEACVIQGVGVTETPLMKED 38 66 RRAVLPQEEEGSGGGQLVTE 14 19 KKWVQDSMKYLDQKSPTPKP 38 67 RRAVLPQEEEGSGGGQLVTE 15 20 SHHSDESDELVTDFPTDLPA 39 68 LNAVNNSLTPQSTKVPSLFE 15 21 SHHSDESDELVTDFPTDLPA 39 69 LNAVNNSLTPQSTKVPSLFE 16 22 DESDELVTDFPTDLPATEVF 40 70 NAVNNSLTPQSTKVPSLFEF 16 23 DESDELVTDFPTDLPATEVF 40 71 NAVNNSLTPQSTKVPSLFEF 17 24 FPTDFPTDLPATEVFTPVVP 41 72 TFVLSALQPSPTHSSSNTQR 17 25 FPTDFPTDLPATEVFTPVVP 41 73 TFVLSALQPSPTHSSSNTQR 18 26 FPTDLPATEVFTPVVPTVDT 42 74 RQGWALRPVLPTQSAHDPPA 18 27 FPTDLPATEVFTPVVPTVDT 42 75 RQGWALRPVLPTQSAHDPPA 19 28 PATEVFTPVVPTVDTYDGRG 43 76 QKKAKNLDAITTPDPTTNAS 19 29 PATEVFTPVVPTVDTYDGRG 43 77 QKKAKNLDAITTPDPTTNAS 20 30 PGAQGLPGVGLTPSAAQTAR 44 78 FLSLSQGQESQTELPNPRIS 20 31 PGAQGLPGVGLTPSAAQTAR 44 79 FLSLSQGQESQTELPNPRIS 21 32 RGETRCEQDRPSPTTAPPAP 45 80 LSLALVTNSAPTSSSTKKTQ 21 33 RGETRCEQDRPSPTTAPPAP 45 81 LSLALVTNSAPTSSSTKKTQ 22 34 ETRCEQDRPSPTTAPPAPPS 46 82 LISPLAQAVRSSSRTPSDKP 22 35 ETRCEQDRPSPTTAPPAPPS 46 83 LISPLAQAVRSSSRTPSDKP 23 36 TRCEQDRPSPTTAPPAPPSP 47 84 DDENTAQFVHVSESFPHPGF 23 37 TRCEQDRPSPTTAPPAPPSP 47 85 DDENTAQFVHVSESFPHPGF 24 38 PSPTTAPPAPPSPSPSPVPK 48 86 SESFPHPGFNMSLLENHTRQ 24 39 PSPTTAPPAPPSPSPSPVPK 48 87 SESFPHPGFNMSLLENHTRQ 25 40 PTTAPPAPPSPSPSPVPKSP 49 88 SGWGSIEPENFSFPDDLQCV 25 41 PTTAPPAPPSPSPSPVPKSP 49 89 SGWGSIEPENFSFPDDLQCV 26 42 TAPPAPPSPSPSPVPKSPSV 50 90 RIQRGPGRAFVTIGKIGNMR 26 43 TAPPAPPSPSPSPVPKSPSV 50 91 RIQRGPGRAFVTIGKIGNMR 27 44 STNEFLCDKDKTSTVAPTIH 51 92 EMSRHSLEQKPTDAPPKVLT 27 45 STNEFLCDKDKTSTVAPTIH 51 93 EMSRHSLEQKPTDAPPKVLT 28 46 NEFLCDKDKTSTVAPTIHTT 52 94 CSESLELEDPSSGLGVTKQD 28 47 NEFLCDKDKTSTVAPTIHTT 52 95 CSESLELEDPSSGLGVTKQD 29 48 CDKDKTSTVAPTIHTTVPSP 53 96 LLEFYLAMPFATPMEAELAR *Indicates attachment site of GalNAc residues to Ser/Thr (bold, underlined).

[0156] Glycopeptides were synthesized with and without N-terminal linker for printing on epoxy or NHS-activated glass slides, respectively. Peptides were designed based on known O-glycoproteins using the algorithm NetOGlyc for selection of O-glycosylation sites (Julenius et al. 2005), which in most cases coincides with experimentally determined O-glycosylation found on isolated proteins, as these glycoproteins have served as a training set for the algorithm. Glycopeptides were designed with a single GalNAc at position 12 to allow optimal exposure of peptide sequence flanking the O-glycan taking into consideration that the glycopeptides will be covalently linked primarily through the N-terminal amino acid. Glycopeptides were synthesized at 100 nmol scale using step-wise blocking by acetylation and printing directly with theoretical excess of 50 fold onto slides.

[0157] In one group, the paired peptide and single GalNAc glycopeptide library was further expanded by on-slide glycosylation with one or more recombinant glycosyltransferases to enhance the number of GalNAc O-glycosylation sites using one or more recombinant polypeptide GalNAc-transferases (e.g., GalNAc-T2, -T3 and -T4) (see, U.S. Pat. Nos. 5,876,716 and 6,465,220) (Bennett et al. 1998; Bennett et al. 1996; White et al. 1995) (see also, FIG. 1). Further expansion was achieved by use of a recombinant sialyltransferase, ST6GalNAc-II, to produce STn glycoforms (structure no. 2, Table I) of the GalNAc glycopeptides. T glycoforms were produced by a recombinant Drosophila core1 β3galactosyltransferase, truncated core3 glycoforms (structure no. 6, Table I) were produced with a recombinant human β3GlcNAc-transferase (Iwai et al. 2002), and non-capped type1-core3 glycoforms (structure no. 9, Table I) were produced using β3Gal-T5 (see, U.S. Pat. No. 7,332,279).

[0158] Recombinant enzymes were expressed in insect cells using the baculo-virus system and used after semi-purification by Ni-chromatography (when HIS-tagged) or by ion exchange chromatographies. Glycosylation was monitored by staining of slides with lectins (HPA, VVA) (Sigma) and monoclonal antibodies to Tn (HBTn1, HBSTn1, HBT1) (Dako, Denmark). On-slide glycosylation was performed as follows: slides were quenched for 1 hr in 50 mM ethanolamine in 100 mM sodium borate pH 8, washed extensively in de-ionized water, and spun dry. Slides were blocked for 1 h with 1% BSA in PBS, pH 7.4, and in some cases with NP40 (1%) to reduce background. Slides were washed in PBS/.05% Tween and enzyme reaction mixtures with BSA and detergent were applied and incubated 1-2 hrs at 37° C. Reaction mixtures for polypeptide GalNAc-transferases included MES buffer 125 mM, pH 7.4, 1% NP40, 1% BSA, 250 μM UDP-GalNAc, 2 mM MnCl₂ and 20 μg/mL enzyme. Reaction mixtures for galactosyltransferases included UDP-Gal, sialyltransferases CMP-NeuAc, GlcNAc-transferases UDP-GlcNAc. Following on-slide glycosylation slides were washed with PBS/Tween and processed with antibodies and lectins as described below.

[0159] It will be clear to one of ordinary skill in the art that on-slide glycosylation serves as a method to screen for additional glycoforms of peptides which may be recognized by auto-antibodies present in patients. Once a particular glycoform of a given peptide is found to react with an autoantibody from a patient, the glycopeptide can be resynthesized in solution using the same enzymes used for on-slide glycosylation and the glycan structure and sites of attachment in the peptide confirmed by mass spectrometry (Tarp et al. 2007). The validated glycopeptide can then be incorporated into diagnostic panels as described throughout the specification and in Example 4.

[0160] FIG. 1 illustrates an example of on-slide glycosylation with the polypeptide GalNAc-transferase, GalNAc-T3, to glycosylate peptides and GalNAc glycopeptides having additional unsubstituted Ser/Thr residues to enhance the number of O-glycans. A library of 96 paired GalNAc-glycopeptides/peptides (20-mers with and without a single GalNAc residue at position 12), as designated in Table II, supra, were printed in triplicates (horizontal) on Scineon 16-well slides by JPT (Germany). In each pair in Table II, the glycopeptide of the pair contains a bold, underlined amino acid residue, indicating the site of attachment of GalNAc. Peptide pairs by peptide number are as follows: 1/2, 3/4, 5/6 . . . etc. The slide was reacted with the anti-Tn lectin HPA (1 μg/ml) without (FIG. 1A) and with (FIG. 1B) prior treatment with recombinant polypeptide GalNAc-T3 enzyme for on-slide GalNAc-glycosylation of available unglycosylated sites on peptides and glycopeptides.

[0161] The HPA lectin did not react with all GalNAc-glycopeptides (e.g., glycopeptide nos. 1-5 and 25-29), which is partly due to failure in synthesis and/or printing at these positions, as well as some restrictions of specificity of the GalNAc-binding lectin. The GalNAc-glycopeptides 1-5 did react with the HPA lectin in other experiments, and it was therefore concluded that there was a print failure. Conversion of peptides to GalNAc-glycopeptides by on-slide reaction with a polypeptide GalNAc-transferase is also expected to be dependent on the substrate specificity of the enzyme used, and, e.g., use of GalNAc-T2 rather than T3 may give a partly different labeling pattern.

[0162] Regardless, it is evident that most of the GalNAc-glycopeptides were labeled with HPA (FIG. 1A) and that most of the corresponding paired peptides were labeled only after GalNAc-T3 on-slide glycosylation (FIG. 1B). While HPA may not be expected to react with all GalNAc-glycopeptides, this lectin and other Tn reactive lectins such as HAA, VVA and DBA, as well as monoclonal anti-Tn antibodies (Dako) provide excellent controls for determining the quality of peptide synthesis and printing.

Example 2

High Through-Put Screening of Glycopeptide Microarrays with Human Serum for Identification of Glycopeptides Recognized by Cancer-Associated Auto-Antibodies

[0163] Sera Origin and Handling:

[0164] A panel of human sera was obtained from CHTN (Cooperative Human Tissue Network) and Asterand Inc., under the guidelines of approved agreements by the providers. Sera obtained were from control individuals ("normal") (n=31) and from newly diagnosed cancer patients (n=147) with pancreatic, breast, colon, lung, prostate or ovary primary cancers. Briefly, for cancer sera, all blood samples were obtained on or near date of diagnosis, and serum was processed immediately, flash frozen and stored at -70° C. until shipment. Serum samples received from the providers as frozen aliquots generally of 1-2 mL were brought to room temperature, vortexed and distributed into 20-100 μL aliquots in closeable Eppendorf tubes (vWR) (pre-labeled), and immediately frozen and stored at -70° C. until use.

[0165] Glycopeptide Microarray Method:

[0166] Glycopeptides (20-mers with and without a single GalNAc residue at position 12) and control structures (corresponding unglycosylated peptides) were printed on Corning (Corning, N.Y.), Scienion (Germany) or Schott Nexterion® Slide H or Schott Nexterion® Slide H MPX 16 (Schott A G, Mainz, Germany) by JPT (Germany). Triplicates or quadruplicates of all compounds were printed at optimal concentrations (1-50 μM) or 50× excess relative to scale of synthesis for spot-synthesized glycopeptides printed without purification. After printing, slides were incubated for 1 hour (h) in a humidified hybridization chamber with 70-100% relative humidity and stored until use at 4° C. Unspotted slide areas were blocked for 1 h with 25 mM ethanolamine in 100 mM sodium borate pH 8.5. If an enzyme step was needed either to increase sites of GaLNac attachments in peptides or extend O-glycans to T, STn, core 3 or other structures, addition of the enzyme reaction mixture in 25-35 μL was made in appropriate wells and the slide incubated at 37° C. for 1 h, after which it was washed in PBS/.05% Tween and then PBS, and then spun dry.

[0167] Human sera (usually diluted 1:25), monoclonal antibodies, lectins and the like were added in 25 μL and the slide left at room temperature for 2 h in a moist, humid chamber after which it was washed as above and spun dry (if no superstructure). Secondary antibodies were added at appropriate concentrations (for human sera, usually 1:1500 for anti-human IgG with a Cy3 chromophore). In some cases, for some controls, lectin-Cy3 was used directly as final step. The final step was washing as above, with a brief de-ionized water wash and the slide spun dry for scanning Analysis was made on a GenePix 4200AL Scanner at PMT 400 and power mode 10-50. Data were analyzed and plotted using Microsoft Excel.

[0168] Identification of Cancer-Associated Auto-Antibodies to GalNAc-Glycopeptide Epitopes:

[0169] Glycopeptides (20-mers with and without a single GalNAc residue at position 12) were printed in triplicates (horizontal) on Scineon 16-well slides by JPT (Germany) and bound human IgG antibodies detected by a labeled secondary anti-human IgG antibody. FIG. 2 illustrates examples of reactivities of serum (1:25 dilution) from a newly diagnosed prostate cancer patient (#762 in Panel A) and a normal control serum from a healthy individual (#174 in Panel B) on the library of 96 paired peptides/GalNAc-glycopeptides as designated in Table II. Candidate cancer-associated IgG antibodies identified in the cancer serum directed to GalNAc glycopeptide epitopes are indicated by open circles labeled 20/21, 62/63, and 86/87 for the paired peptide and GalNAc glycopeptides, respectively. The analysis demonstrates that serum of cancer patient contain IgG antibodies specifically reacting with epitopes found on GalNAc glycopeptides 21, 63, and 87, and since these antibodies do not react with the corresponding unglycosylated peptides 20, 62, and 86, it may be concluded that the epitopes are comprised of a GalNAc-peptide epitope including both the O-glycan part as well as part of the peptide sequence.

[0170] Human IgG antibodies from several cancers were shown to bind selectively to several GalNAc-glycopeptides and not the corresponding peptide, and such antibodies were not detected in healthy individuals. An example of this is shown in FIG. 2, where a prostate cancer serum (prca#762) labels three GalNAc-glycopeptides (#21, 63, and 87), and not the corresponding peptide (#20, 62, and 86). These three candidates were also identified in other cancer sera but not in the controls as shown in FIGS. 3-5, and described, infra.

[0171] FIG. 3 illustrates results of screening 147 cancer sera and 31 control sera (normal sera) on the glycopeptide pair #20/21. Sera obtained from newly diagnosed cancer patients with lung, colon, ovary, prostate, pancreas or breast tumors as well as controls (as indicated) were reacted with the 96-peptide array on Scineon 16-well slides (1:25 dilution) followed by cy3 labeled anti-human IgG (diluted to 1:1500). Arrays were analyzed on GenePix 4200 scanner at 400 pmt with 50 power and relative intensities graphed.

[0172] FIG. 4 illustrates results of screening 147 cancer sera and 31 normal sera on the glycopeptide pair #62/63. FIG. 5 illustrates results of screening 147 cancer sera and 31 normal sera on the glycopeptide pair #86/87.

[0173] Peptides 20/21, each having the sequence identified as SEQ ID NO: 15, are derived from SPP1 (Ensg00000118785) GenBank Accession No. NP_001035147 (SEQ ID NO: 61), peptides 62/63, each having the sequence identified as SEQ ID NO: 36 are derived from LAMP2 (Ensg00000005893) (ENST00000200639) protein (ENSP00000200639) GenBank Accession No. NP_002285 (SEQ ID NO: 64), and peptides 86/87, each having the sequence identified as SEQ ID NO: 49, are derived from KLK1 (gene: Ensg00000167748) (transcript: ENST00000301420) (protein: ENSP00000301420) (GenBank Accession No. NP_002248) (SEQ ID NO: 66). These proteins have broad expression patterns in tissues and hence may induce antibodies in many cancers.

[0174] The presented method using a limited glycopeptide library identified three candidate targets for human disease-associated auto-antibodies. These auto-antibody targets were defined by the discriminating factor that cancer-associated IgG antibodies reacted selectively with a glycopeptide and not the corresponding peptide or other glycopeptides with the same O-glycan or the O-glycan presented as a hapten on an artificial carrier.

Example 3

Generation of Antibodies to Identified Auto-Antibody Targets for Diagnostic Use

[0175] Monoclonal or polyclonal antibodies may be generated to the identified glycopeptide antigens by known methods (Takeuchi et al. 2002; Hanisch et al. 1995; Reis et al. 1998; Sorensen et al. 2006). Briefly, the following procedure serves as an example for generation of antibodies with the desired glycopeptides specificity but other procedures leading to the same result will be known to the skilled in the art.

[0176] Immunization protocol: Glycopeptides are coupled to keyhole limpet hemocyanin (KLH) (Pierce, Rockford, Ill.) using glutaraldehyde. Efficiency of conjugation is assessed by analyzing the reaction by size exclusion chromatography on a PD-10 column, where the conjugate/glycopeptides ratio can be determined by ELISA using appropriate reagents such as antibodies and lectins detecting the glycopeptides. Essentially all reactivity with a Tn reactive lectin (HPA) is found with the excluded fraction and insignificant reactivity in the included fractions expected to contain peptides. Titration analysis of the KLH conjugate with the corresponding glycopeptide in ELISA indicated conjugation ratio KLH to glycopeptide of approximately 1:200. Female Balb/c (Jackson Labs) wild type mice are injected subcutaneously with 10 or 15 μg of glycopeptide-KLH in a total volume of 200 μl (1:1 mix with Freunds adjuvant, Sigma). Mice received four immunizations 2-4 wks apart, and blood samples are obtained by tail or eye bleeding 1 wk following the third and fourth immunization.

[0177] Hybridoma Production:

[0178] Mouse hybridomas are produced by fusion of splenocytes to NS-1 followed by selection in HAT/HT (Hypoxanthine, Aminopterin, Thymidine). Hybridomas are selected by initial screening by ELISA with GalNAc-glycopeptides and corresponding peptides as well as irrelevant control compounds. Further characterization is done on glycopeptides microarrays as well as on a panel of human cancer cell lines expressing the corresponding aberrant glycoprotein.

[0179] Enzyme-linked immunosorbent assays (ELISA) are performed using 96-well MaxiSorp® plates (Nunc). Plates are coated overnight at 4° C. with 1 μg/ml of glycopeptides in bicarbonate-carbonate buffer (pH 9.5), blocked with 5% BSA in PBS, followed by incubation with sera (diluted in PBS) or monoclonal antibodies for 2 hours (hrs) at room temperature. Bound antibodies are detected with peroxidase-conjugated rabbit anti-mouse immunoglobulins (Dako, Denmark) or isotype specific antibodies peroxidase-conjugated goat anti-mouse IgM, IgG1, IgG2a, IgG2b, or IgG3 (Southern Biotechnology Associates, USA). Plates are developed with o-phenylenediamine tablets (Dako, Denmark) and read at 492 nm. Control antibodies included anti-carbohydrate antibodies HBTn-1 (Tn) and HBSTn-1 (STn) (Dako) and lectins HPA, VVA and HAA (Sigma). Control sera included mice immunized with irrelevant peptides linked to KLH. Human cancer cell lines from different cancers are all obtained from ATCC and cultured as recommended by ATCC.

[0180] Immunocytochemical staining of cells are performed as follows: cells are harvested by trypsination, washed and plated onto multi-well glass slides and fixed for 10 minutes (min) in ice cold acetone or in methanol acetone. Fixed cells are incubated overnight at 4° C. with mouse sera (1:200/1:400/1:800) or hybridoma antibodies, followed by incubation for 45 min at room temperature with FITC-conjugated rabbit anti-mouse immunoglobulins (Dako, Denmark). Slides are mounted in glycerol containing p-phenylenediamine and examined in a fluorescence microscope.

[0181] For further determination of expression of glycopeptides epitope in human cancers immunohistochemistry of fixed and frozen tissue samples are performed. Frozen sections are fixed for 10 min in cold methanol/acetone (50:50). Formalin fixed, paraffin wax embedded tissues of different carcinoma and healthy tissues are obtained from Origine (US) and stained by immunofluorescence or peroxidase techniques. Paraffin sections are dewaxed, rehydrated, and treated with 0.5% H₂O₂ in methanol for 30 min. Section are rinsed in TBS and incubated for 20 min with rabbit nonimmune serum. Sections are rinsed and incubated overnight at 4° C. with primary antibody. Sections are rinsed and incubated with biotin-labeled rabbit anti-mouse serum (Dako, Denmark) diluted 1:200 in TBS for 30 min, rinsed with TBS, and incubated for 1 h with avidin-biotin-peroxidase complex (Dako, Denmark). Sections are rinsed with TBS and developed with 0.05% 3,3'-diaminobenzidine tetrahydrochloride freshly prepared in 0.05 M TBS containing 0.1% H₂O₂. Sections are stained with hematoxylin, dehydrated and mounted.

Example 4

Broad Discovery of O-Glycopeptide Epitopes

[0182] A larger library of GalNAc O-glycopeptides designed with GalNAc linked to serine or threonine at position 12 in 20-mer peptides was produced as described in Example 1. Peptides were selected among cell membrane and secreted human proteins that contain serine and threonine residues that are predicted to be O-glycosylated by the NetOGlyc algorithm (Julenius et al. 2005). A total of 960 GalNAc glycopeptides were synthesized at 0.5 mg scale (Sigma) and printed on microarray hydrogel slides (Schott Nexterion) as described in Example 1. Slides were reacted with human cancer sera (ovarian, breast, colon, lung, pancreas, prostate, 235 total sera) and control sera (145 healthy, 20 inflammatory, 20 benign tumor). Glycopeptides preferentially reactive with human IgG antibodies from cancer patients compared to controls were selected for resynthesis at 5 mg scale with amidated C-terminus (New England Peptide (Gardner, Mass.) (Table III). Resynthesized glycopeptides were reprinted on microarray slides at 50 μM and reacted with human cancer sera (Total 286 sera: 32 ovarian cancer (O), 38 breast cancer (B), 54 colon cancer (C), 17 lung cancer (L), and 145 age and sex matched control normal sera as described in Example 1. Table III lists glycopeptides exclusively or preferentially reactive with human IgG from cancer patients compared to controls and examples of reactivities are shown in FIG. 6.

TABLE-US-00003 TABLE III Glycopeptides reactive with IgG antibodies from human cancer sera Peptide Sequence number ID Protein Sequence 166 82 Oncostatin H2N-TKAGRGASQPPTPTPASDAF-amide 113 83 gp95 H2N-EEEPEETAEDTTEDTEQDED-amide 218 84 MUC13 H2N-TASTTANTPFPTATSPAPPI-amide 225 85 MUC13 H2N-PAPPIISTHSSSTIPTPAPP-amide 275a 86 Ceruplasm H2N-LAKMYYSAVEPTKDIFTGLI-amide 284 87 CD46 H2N-PSSTKPPALSHSVSTSSTTK-amide 308 88 EBAG9 H2N-LEPDYFKDMTPTIRKTQKIV-amide 318a 89 MUC17 H2N-STMPVVSSEASTHSTTPVDT-amide 324 90 MUC17 H2N-STHSTTPVDTSTPVTTSTEA-amide 370 91 IL6-R H2N-IPPEDTASTRSSFTVQDLKP-amide 382 92 R-PTP-alpha H2N-EAKTSNPTSSLTSLSVAPTF-amide 389 93 R-PTP-alpha H2N-ARTEPWEGNSSTAATTPETF-amide 443 94 ODAM H2N-VDPLQLQTPPQTQPGPSHVM-amide 450 95 ODAM H2N-SPKPSTTNVFTSAVDQTITP-amide 456 96 IGFB-3 H2N-PAPPAPGNASESEEDRSAGS-amide 458 97 IGFB-3 H2N-ASESEEDRSAGSVESPSVSS-amide 465 98 MUC15 H2N-ANLNSDKENITTSNLKASHS-amide 484 99 MUC15 H2N-LTTNSDSFTGFTPYQEKTTL-amide 485 100 MUC15 H2N-SFTGFTPYQEKTTLQPTLKF-amide 485a 101 MUC15 H2N-SFTGFTPYQEKTTLQPTLKF-amide 505 102 TPBG Trophob H2N-SPTSSASSFSSSAPFLASAV-amide 522 103 IgA1 hinge H2N-TVPCPVPSTPPTPSPSTPPT-amide 524 104 IgA1 hinge H2N-VPSTPPTPSPSTPPTPSPSC-amide 544 105 R-PTP-N H2N-SEPPKAARPPVTPVLLEKKS-amide 546 106 R-PTP-N H2N-GQSQPTVAGQPSARPAAEEY-amide 558 107 CMRF35 H2N-TPASITAAKTSTITTAFPPV-amide 569 108 TNF-RSF1B H2N-GNASMDAVCTSTSPTRSMAP-amide 585 109 CGB2 H2N-TDCGGPKDHPLTCDDPRFQA-amide 587 110 CGB2 H2N-AQASSSSKAPPPSLPSPSRLP-amide 592 111 Acrosomal SP H2N-PLSELESGEQPSDEQPSGEH-amide 601 112 Cadherin 1 H2N-PQRSSTAILQVSVTDTNDNH-amide (CD) 605 113 CD Ovomorolin H2N-GALPGTSVMEVTATDADDDV-amide 612 114 CD Mucin like H2N-EQEPPSTDVPPSPEAGGTTG-amide 676 115 Inhibin alpha H2N-RPEATPFLVAHTRTRPPSGG-amide 690 116 IGF-BP-6 H2N-PGTSTTPSQPNSAGVQDTEM-amide 739 117 LTBP1 H2N-EVAPEASTSSASQVIAPTQV-amide 752 118 CD 248 H2N-QPPDFALAYRPSFPEDREPQ-amide Endosialin 755 119 CD 248 H2N-LSVTRPVVVSATHPTLPSAH-amide Endosialin 757 120 CD 248 H2N-PSAHQPPVIPATHPALSRDH-amide Endosialin 765 121 CD 248 H2N-APDALVLRTQATQLPIIPTA-amide Endosialin 823 122 ICAM-1 H2N-GALFPGPGNAQTSVSPSKVI-amide 827 123 ICAM-1 H2N-HLALGDQRLNPTVTYGNDSF-amide 827a 124 ICAM-1 H2N-HLALGDQRLNPTVTYGNDSF-amide 848 125 MUC1 H2N-PATEPASGSAATWGQDVTSV-amide 852 126 MUC1 H2N-VPVTRPALGSTTPPAHDVTS-amide 859 127 MUC1 H2N-NVTSASGSASGSASTLVHNG-amide 859a 128 MUC1 H2N-NVTSASGSASGSASTLVHNG-amide 863 129 MUC1 H2N-GTSARATTTPASKSTPFSIP-amide 863a 130 MUC1 H2N-GTSARATTTPASKSTPFSIP-amide 870 131 MUC1 H2N-SDTPTTLASHSTKTDASSTH-amide 873 132 MUC1 H2N-TKTDASSTHHSTVPPLTSSN-amide 883 133 MUC1 H2N-TDYYQELQRDISEMFLQIYK-amide 889 134 MUC1 H2N-HDVETQFNQYKTEAASRYNL-amide 893 135 MUC1 H2N-ASRYNLTISDVSVSDVPFPF-amide 894 136 MUC1 H2N-RYNLTISDVSVSDVPFPFSA-amide 895 137 MUC1 H2N-DVSVSDVPFPFSAQSGAGVP-amide 914 138 MUC4 H2N-TAGRPTGQSSPTSPSASPQE-amide 931 139 MUC4 H2N-SLASQATDIFSTVPPTPPSI-amide 934 140 MUC4 H2N-FSTVPPTPPSITSTGLTSPQ-amide 936 141 MUC4 H2N-PTPPSITSTGLTSPQTETHT-amide 941 142 MUC4 H2N-LTSPQTETHTLSPSGSGKTF-amide 977 143 MUC4 H2N-TDTSSASTGHATPLPVTSLS-amide 983 144 MUC4 H2N-HATPLAVSSATSASTVSSDS-amide 852-C3 145 MUC1 H2N- VPVTRPALGSTT[GlcNAc]PPAHDVTS- amide 931-C3 146 MUC4 H2N- SLASQATDIFST[GlcNAc]VPPTPPSI- amide *Bold S or T indicates attachment site of GalNAc residues to Ser/Thr; Bold [GlcNAc] indicates an addition of glucosamine attached to GalNAc (in bold). Bold, underlined name indicates peptide assayed in graphs in FIG. 6.

[0183] Since IgG antibodies reactive with glycopeptides epitopes recognize both the peptide sequence in close proximity to the O-glycan (i.e., one to five or even eighth residues N- and C-terminal of the O-glycan) as well as the O-glycan structure, it is clear that different glycoforms of one glycopeptide may be recognized by different IgG antibodies and these antibodies may be found in different cancer patients. In this Example, GalNAc-glycopeptides from MUC1 and MUC4 were also produced as the core 3 GlcNAcβ1-3GalNAcα1-O-Ser/Thr glycoform with a recombinant core 3 synthase as described in Example 1. Two core 3 glycopeptides (#852-C3 and #931-C3) were identified as exclusively recognized by IgG antibodies in sera from cancer patients.

[0184] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0185] It is further to be understood that all values are approximate, and are provided for description.

[0186] Patents, patent applications, publications, product descriptions, and protocols are cited throughout this application, the disclosures of which are incorporated herein by reference in their entireties for all purposes.

LITERATURE CITED

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Sequence CWU 1

1

1461273PRTHomo sapiens 1Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr 1 5 10 15 Val Leu Thr Val Val Thr Gly Ser Gly His Ala Ser Ser Thr Pro Gly 20 25 30 Gly Glu Lys Glu Thr Ser Ala Thr Gln Arg Ser Ser Val Pro Ser Ser 35 40 45 Thr Glu Lys Asn Ala Leu Ser Thr Gly Val Ser Phe Phe Phe Leu Ser 50 55 60 Phe His Ile Ser Asn Leu Gln Phe Asn Ser Ser Leu Glu Asp Pro Ser 65 70 75 80 Thr Asp Tyr Tyr Gln Glu Leu Gln Arg Asp Ile Ser Glu Met Phe Leu 85 90 95 Gln Ile Tyr Lys Gln Gly Gly Phe Leu Gly Leu Ser Asn Ile Lys Phe 100 105 110 Arg Pro Gly Ser Val Val Val Gln Leu Thr Leu Ala Phe Arg Glu Gly 115 120 125 Thr Ile Asn Val His Asp Val Glu Thr Gln Phe Asn Gln Tyr Lys Thr 130 135 140 Glu Ala Ala Ser Arg Tyr Asn Leu Thr Ile Ser Asp Val Ser Val Ser 145 150 155 160 Asp Val Pro Phe Pro Phe Ser Ala Gln Ser Gly Ala Gly Val Pro Gly 165 170 175 Trp Gly Ile Ala Leu Leu Val Leu Val Cys Val Leu Val Ala Leu Ala 180 185 190 Ile Val Tyr Leu Ile Ala Leu Ala Val Cys Gln Cys Arg Arg Lys Asn 195 200 205 Tyr Gly Gln Leu Asp Ile Phe Pro Ala Arg Asp Thr Tyr His Pro Met 210 215 220 Ser Glu Tyr Pro Thr Tyr His Thr His Gly Arg Tyr Val Pro Pro Ser 225 230 235 240 Ser Thr Asp Arg Ser Pro Tyr Glu Lys Val Ser Ala Gly Asn Gly Gly 245 250 255 Ser Ser Leu Ser Tyr Thr Asn Pro Ala Val Ala Ala Thr Ser Ala Asn 260 265 270 Leu 2264PRTHomo sapiens 2Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr 1 5 10 15 Val Leu Thr Ala Thr Thr Ala Pro Lys Pro Ala Thr Val Val Thr Gly 20 25 30 Ser Gly His Ala Ser Ser Thr Pro Gly Gly Glu Lys Glu Thr Ser Ala 35 40 45 Thr Gln Arg Ser Ser Val Pro Ser Ser Thr Glu Lys Asn Ala Phe Asn 50 55 60 Ser Ser Leu Glu Asp Pro Ser Thr Asp Tyr Tyr Gln Glu Leu Gln Arg 65 70 75 80 Asp Ile Ser Glu Met Phe Leu Gln Ile Tyr Lys Gln Gly Gly Phe Leu 85 90 95 Gly Leu Ser Asn Ile Lys Phe Arg Pro Gly Ser Val Val Val Gln Leu 100 105 110 Thr Leu Ala Phe Arg Glu Gly Thr Ile Asn Val His Asp Val Glu Thr 115 120 125 Gln Phe Asn Gln Tyr Lys Thr Glu Ala Ala Ser Arg Tyr Asn Leu Thr 130 135 140 Ile Ser Asp Val Ser Val Ser Asp Val Pro Phe Pro Phe Ser Ala Gln 145 150 155 160 Ser Gly Ala Gly Val Pro Gly Trp Gly Ile Ala Leu Leu Val Leu Val 165 170 175 Cys Val Leu Val Ala Leu Ala Ile Val Tyr Leu Ile Ala Leu Ala Val 180 185 190 Cys Gln Cys Arg Arg Lys Asn Tyr Gly Gln Leu Asp Ile Phe Pro Ala 195 200 205 Arg Asp Thr Tyr His Pro Met Ser Glu Tyr Pro Thr Tyr His Thr His 210 215 220 Gly Arg Tyr Val Pro Pro Ser Ser Thr Asp Arg Ser Pro Tyr Glu Lys 225 230 235 240 Val Ser Ala Gly Asn Gly Gly Ser Ser Leu Ser Tyr Thr Asn Pro Ala 245 250 255 Val Ala Ala Thr Ser Ala Asn Leu 260 3255PRTHomo sapiens 3Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr 1 5 10 15 Val Leu Thr Val Val Thr Gly Ser Gly His Ala Ser Ser Thr Pro Gly 20 25 30 Gly Glu Lys Glu Thr Ser Ala Thr Gln Arg Ser Ser Val Pro Ser Ser 35 40 45 Thr Glu Lys Asn Ala Phe Asn Ser Ser Leu Glu Asp Pro Ser Thr Asp 50 55 60 Tyr Tyr Gln Glu Leu Gln Arg Asp Ile Ser Glu Met Phe Leu Gln Ile 65 70 75 80 Tyr Lys Gln Gly Gly Phe Leu Gly Leu Ser Asn Ile Lys Phe Arg Pro 85 90 95 Gly Ser Val Val Val Gln Leu Thr Leu Ala Phe Arg Glu Gly Thr Ile 100 105 110 Asn Val His Asp Val Glu Thr Gln Phe Asn Gln Tyr Lys Thr Glu Ala 115 120 125 Ala Ser Arg Tyr Asn Leu Thr Ile Ser Asp Val Ser Val Ser Asp Val 130 135 140 Pro Phe Pro Phe Ser Ala Gln Ser Gly Ala Gly Val Pro Gly Trp Gly 145 150 155 160 Ile Ala Leu Leu Val Leu Val Cys Val Leu Val Ala Leu Ala Ile Val 165 170 175 Tyr Leu Ile Ala Leu Ala Val Cys Gln Cys Arg Arg Lys Asn Tyr Gly 180 185 190 Gln Leu Asp Ile Phe Pro Ala Arg Asp Thr Tyr His Pro Met Ser Glu 195 200 205 Tyr Pro Thr Tyr His Thr His Gly Arg Tyr Val Pro Pro Ser Ser Thr 210 215 220 Asp Arg Ser Pro Tyr Glu Lys Val Ser Ala Gly Asn Gly Gly Ser Ser 225 230 235 240 Leu Ser Tyr Thr Asn Pro Ala Val Ala Ala Thr Ser Ala Asn Leu 245 250 255 420PRTArtificial SequenceSynthetic peptides 4Thr Ser Ala Pro Asp Thr Arg Pro Ala Pro Gly Ser Thr Ala Pro Pro 1 5 10 15 Ala His Gly Val 20 520PRTArtificial SequenceSynthetic peptides 5Ser Ala Pro Asp Thr Arg Pro Ala Pro Gly Ser Thr Ala Pro Pro Ala 1 5 10 15 His Gly Val Thr 20 620PRTArtificial SequenceSynthetic peptides 6Pro Gly Ser Thr Ala Pro Pro Ala His Gly Val Thr Ser Ala Pro Asp 1 5 10 15 Thr Arg Pro Ala 20 720PRTArtificial SequenceSynthetic peptides 7Gly Ser Thr Ala Pro Pro Ala His Gly Val Thr Ser Ala Pro Asp Thr 1 5 10 15 Arg Pro Ala Pro 20 820PRTArtificial SequenceSynthetic peptides 8Pro Pro Ala His Gly Val Thr Ser Ala Pro Asp Thr Arg Pro Ala Pro 1 5 10 15 Gly Ser Thr Ala 20 920PRTArtificial SequenceSynthetic peptides 9Phe Tyr Leu Ala Met Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala 1 5 10 15 Arg Arg Ser Leu 20 1020PRTArtificial SequenceSynthetic peptides 10Lys Phe Ser Glu Phe Trp Asp Leu Asp Pro Glu Val Arg Pro Thr Ser 1 5 10 15 Ala Val Ala Ala 20 1120PRTArtificial SequenceSynthetic peptides 11Pro Leu Val Glu Gln Gly Arg Val Arg Ala Ala Thr Val Gly Ser Leu 1 5 10 15 Ala Gly Gln Pro 20 1220PRTArtificial SequenceSynthetic peptides 12Leu Trp Ser Leu Cys Trp Ser Leu Ala Ile Ala Thr Pro Leu Pro Pro 1 5 10 15 Thr Ser Ala His 20 1320PRTArtificial SequenceSynthetic peptides 13Leu Glu Ala Cys Val Ile Gln Gly Val Gly Val Thr Glu Thr Pro Leu 1 5 10 15 Met Lys Glu Asp 20 1420PRTArtificial SequenceSynthetic peptides 14Lys Lys Trp Val Gln Asp Ser Met Lys Tyr Leu Asp Gln Lys Ser Pro 1 5 10 15 Thr Pro Lys Pro 20 1520PRTArtificial SequenceSynthetic peptides 15Ser His His Ser Asp Glu Ser Asp Glu Leu Val Thr Asp Phe Pro Thr 1 5 10 15 Asp Leu Pro Ala 20 1620PRTArtificial SequenceSynthetic peptides 16Asp Glu Ser Asp Glu Leu Val Thr Asp Phe Pro Thr Asp Leu Pro Ala 1 5 10 15 Thr Glu Val Phe 20 1720PRTArtificial SequenceSynthetic peptides 17Phe Pro Thr Asp Phe Pro Thr Asp Leu Pro Ala Thr Glu Val Phe Thr 1 5 10 15 Pro Val Val Pro 20 1820PRTArtificial SequenceSynthetic peptides 18Phe Pro Thr Asp Leu Pro Ala Thr Glu Val Phe Thr Pro Val Val Pro 1 5 10 15 Thr Val Asp Thr 20 1920PRTArtificial SequenceSynthetic peptides 19Pro Ala Thr Glu Val Phe Thr Pro Val Val Pro Thr Val Asp Thr Tyr 1 5 10 15 Asp Gly Arg Gly 20 2020PRTArtificial SequenceSynthetic peptides 20Pro Gly Ala Gln Gly Leu Pro Gly Val Gly Leu Thr Pro Ser Ala Ala 1 5 10 15 Gln Thr Ala Arg 20 2120PRTArtificial SequenceSynthetic peptides 21Arg Gly Glu Thr Arg Cys Glu Gln Asp Arg Pro Ser Pro Thr Thr Ala 1 5 10 15 Pro Pro Ala Pro 20 2220PRTArtificial SequenceSynthetic peptides 22Glu Thr Arg Cys Glu Gln Asp Arg Pro Ser Pro Thr Thr Ala Pro Pro 1 5 10 15 Ala Pro Pro Ser 20 2320PRTArtificial SequenceSynthetic peptides 23Thr Arg Cys Glu Gln Asp Arg Pro Ser Pro Thr Thr Ala Pro Pro Ala 1 5 10 15 Pro Pro Ser Pro 20 2420PRTArtificial SequenceSynthetic peptides 24Pro Ser Pro Thr Thr Ala Pro Pro Ala Pro Pro Ser Pro Ser Pro Ser 1 5 10 15 Pro Val Pro Lys 20 2520PRTArtificial SequenceSynthetic peptides 25Pro Thr Thr Ala Pro Pro Ala Pro Pro Ser Pro Ser Pro Ser Pro Val 1 5 10 15 Pro Lys Ser Pro 20 2620PRTArtificial SequenceSynthetic peptides 26Thr Ala Pro Pro Ala Pro Pro Ser Pro Ser Pro Ser Pro Val Pro Lys 1 5 10 15 Ser Pro Ser Val 20 2720PRTArtificial SequenceSynthetic peptides 27Ser Thr Asn Glu Phe Leu Cys Asp Lys Asp Lys Thr Ser Thr Val Ala 1 5 10 15 Pro Thr Ile His 20 2820PRTArtificial SequenceSynthetic peptides 28Asn Glu Phe Leu Cys Asp Lys Asp Lys Thr Ser Thr Val Ala Pro Thr 1 5 10 15 Ile His Thr Thr 20 2920PRTArtificial SequenceSynthetic peptides 29Cys Asp Lys Asp Lys Thr Ser Thr Val Ala Pro Thr Ile His Thr Thr 1 5 10 15 Val Pro Ser Pro 20 3020PRTArtificial SequenceSynthetic peptides 30Asp Lys Thr Ser Thr Val Ala Pro Thr Ile His Thr Thr Val Pro Ser 1 5 10 15 Pro Thr Thr Thr 20 3120PRTArtificial SequenceSynthetic peptides 31Lys Thr Ser Thr Val Ala Pro Thr Ile His Thr Thr Val Pro Ser Pro 1 5 10 15 Thr Thr Thr Pro 20 3220PRTArtificial SequenceSynthetic peptides 32Thr Val Ala Pro Thr Ile His Thr Thr Val Pro Ser Pro Thr Thr Thr 1 5 10 15 Pro Thr Pro Lys 20 3320PRTArtificial SequenceSynthetic peptides 33Ala Pro Thr Ile His Thr Thr Val Pro Ser Pro Thr Thr Thr Pro Thr 1 5 10 15 Pro Lys Glu Lys 20 3420PRTArtificial SequenceSynthetic peptides 34Pro Thr Ile His Thr Thr Val Pro Ser Pro Thr Thr Thr Pro Thr Pro 1 5 10 15 Lys Glu Lys Pro 20 3520PRTArtificial SequenceSynthetic peptides 35Thr Ile His Thr Thr Val Pro Ser Pro Thr Thr Thr Pro Thr Pro Lys 1 5 10 15 Glu Lys Pro Glu 20 3620PRTArtificial SequenceSynthetic peptides 36Thr Pro Thr Pro Lys Glu Lys Pro Glu Ala Gly Thr Tyr Ser Val Asn 1 5 10 15 Asn Gly Asn Asp 20 3720PRTArtificial SequenceSynthetic peptides 37Ala Cys Leu Ala Val Ser Ala Gly Pro Val Pro Thr Pro Pro Asp Asn 1 5 10 15 Ile Gln Val Gln 20 3820PRTArtificial SequenceSynthetic peptides 38Arg Arg Ala Val Leu Pro Gln Glu Glu Glu Gly Ser Gly Gly Gly Gln 1 5 10 15 Leu Val Thr Glu 20 3920PRTArtificial SequenceSynthetic peptides 39Leu Asn Ala Val Asn Asn Ser Leu Thr Pro Gln Ser Thr Lys Val Pro 1 5 10 15 Ser Leu Phe Glu 20 4020PRTArtificial SequenceSynthetic peptides 40Asn Ala Val Asn Asn Ser Leu Thr Pro Gln Ser Thr Lys Val Pro Ser 1 5 10 15 Leu Phe Glu Phe 20 4120PRTArtificial SequenceSynthetic peptides 41Thr Phe Val Leu Ser Ala Leu Gln Pro Ser Pro Thr His Ser Ser Ser 1 5 10 15 Asn Thr Gln Arg 20 4220PRTArtificial SequenceSynthetic peptides 42Arg Gln Gly Trp Ala Leu Arg Pro Val Leu Pro Thr Gln Ser Ala His 1 5 10 15 Asp Pro Pro Ala 20 4320PRTArtificial SequenceSynthetic peptides 43Gln Lys Lys Ala Lys Asn Leu Asp Ala Ile Thr Thr Pro Asp Pro Thr 1 5 10 15 Thr Asn Ala Ser 20 4420PRTArtificial SequenceSynthetic peptides 44Phe Leu Ser Leu Ser Gln Gly Gln Glu Ser Gln Thr Glu Leu Pro Asn 1 5 10 15 Pro Arg Ile Ser 20 4520PRTArtificial SequenceSynthetic peptides 45Leu Ser Leu Ala Leu Val Thr Asn Ser Ala Pro Thr Ser Ser Ser Thr 1 5 10 15 Lys Lys Thr Gln 20 4620PRTArtificial SequenceSynthetic peptides 46Leu Ile Ser Pro Leu Ala Gln Ala Val Arg Ser Ser Ser Arg Thr Pro 1 5 10 15 Ser Asp Lys Pro 20 4720PRTArtificial SequenceSynthetic peptides 47Asp Asp Glu Asn Thr Ala Gln Phe Val His Val Ser Glu Ser Phe Pro 1 5 10 15 His Pro Gly Phe 20 4820PRTArtificial SequenceSynthetic peptides 48Ser Glu Ser Phe Pro His Pro Gly Phe Asn Met Ser Leu Leu Glu Asn 1 5 10 15 His Thr Arg Gln 20 4920PRTArtificial SequenceSynthetic peptides 49Ser Gly Trp Gly Ser Ile Glu Pro Glu Asn Phe Ser Phe Pro Asp Asp 1 5 10 15 Leu Gln Cys Val 20 5020PRTArtificial SequenceSynthetic peptides 50Arg Ile Gln Arg Gly Pro Gly Arg Ala Phe Val Thr Ile Gly Lys Ile 1 5 10 15 Gly Asn Met Arg 20 5120PRTArtificial SequenceSynthetic peptides 51Glu Met Ser Arg His Ser Leu Glu Gln Lys Pro Thr Asp Ala Pro Pro 1 5 10 15 Lys Val Leu Thr 20 5220PRTArtificial SequenceSynthetic peptides 52Cys Ser Glu Ser Leu Glu Leu Glu Asp Pro Ser Ser Gly Leu Gly Val 1 5 10 15 Thr Lys Gln Asp 20 5320PRTArtificial SequenceSynthetic peptides 53Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe Ala Thr Pro Met Glu Ala 1 5 10 15 Glu Leu Ala Arg 20 54203PRTHomo sapiens 54Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr 1 5 10 15 Val Leu Thr Val Val Thr Gly Ser Gly His Ala Ser Ser Thr Pro Gly 20 25 30 Gly Glu Lys Glu Thr Ser Ala Thr Gln Arg Ser Ser Val Pro Ser Ser 35 40 45 Thr Glu Lys Asn Ala Ile Pro Ala Pro Thr Thr Thr Lys Ser Cys Arg 50 55 60 Glu Thr Phe Leu Lys Cys Phe Cys Arg Phe Ile Asn Lys Gly Val Phe 65 70 75 80 Trp Ala Ser Pro Ile Leu Ser Ser Val Ser Asp Val Pro Phe Pro Phe 85 90 95 Ser Ala Gln Ser Gly Ala Gly Val Pro Gly Trp Gly Ile Ala Leu Leu 100 105 110 Val Leu Val Cys Val Leu Val Ala Leu Ala Ile Val Tyr Leu Ile Ala 115 120

125 Leu Ala Val Cys Gln Cys Arg Arg Lys Asn Tyr Gly Gln Leu Asp Ile 130 135 140 Phe Pro Ala Arg Asp Thr Tyr His Pro Met Ser Glu Tyr Pro Thr Tyr 145 150 155 160 His Thr His Gly Arg Tyr Val Pro Pro Ser Ser Thr Asp Arg Ser Pro 165 170 175 Tyr Glu Lys Val Ser Ala Gly Asn Gly Gly Ser Ser Leu Ser Tyr Thr 180 185 190 Asn Pro Ala Val Ala Ala Thr Ser Ala Asn Leu 195 200 55150PRTHomo sapiens 55Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr 1 5 10 15 Val Leu Thr Val Val Thr Gly Ser Gly His Ala Ser Ser Thr Pro Gly 20 25 30 Gly Glu Lys Glu Thr Ser Ala Thr Gln Arg Ser Ser Val Pro Ser Ser 35 40 45 Thr Glu Lys Asn Ala Phe Asn Ser Ser Leu Glu Asp Pro Ser Thr Asp 50 55 60 Tyr Tyr Gln Glu Leu Gln Arg Asp Ile Ser Glu Met Ala Val Cys Gln 65 70 75 80 Cys Arg Arg Lys Asn Tyr Gly Gln Leu Asp Ile Phe Pro Ala Arg Asp 85 90 95 Thr Tyr His Pro Met Ser Glu Tyr Pro Thr Tyr His Thr His Gly Arg 100 105 110 Tyr Val Pro Pro Ser Ser Thr Asp Arg Ser Pro Tyr Glu Lys Val Ser 115 120 125 Ala Gly Asn Gly Gly Ser Ser Leu Ser Tyr Thr Asn Pro Ala Val Ala 130 135 140 Ala Thr Ser Ala Asn Leu 145 150 56159PRTHomo sapiens 56Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr 1 5 10 15 Val Leu Thr Ala Thr Thr Ala Pro Lys Pro Ala Thr Val Val Thr Gly 20 25 30 Ser Gly His Ala Ser Ser Thr Pro Gly Gly Glu Lys Glu Thr Ser Ala 35 40 45 Thr Gln Arg Ser Ser Val Pro Ser Ser Thr Glu Lys Asn Ala Phe Asn 50 55 60 Ser Ser Leu Glu Asp Pro Ser Thr Asp Tyr Tyr Gln Glu Leu Gln Arg 65 70 75 80 Asp Ile Ser Glu Met Ala Val Cys Gln Cys Arg Arg Lys Asn Tyr Gly 85 90 95 Gln Leu Asp Ile Phe Pro Ala Arg Asp Thr Tyr His Pro Met Ser Glu 100 105 110 Tyr Pro Thr Tyr His Thr His Gly Arg Tyr Val Pro Pro Ser Ser Thr 115 120 125 Asp Arg Ser Pro Tyr Glu Lys Val Ser Ala Gly Asn Gly Gly Ser Ser 130 135 140 Leu Ser Tyr Thr Asn Pro Ala Val Ala Ala Thr Ser Ala Asn Leu 145 150 155 5714507PRTHomo sapiens 57Met Leu Lys Pro Ser Gly Leu Pro Gly Ser Ser Ser Pro Thr Arg Ser 1 5 10 15 Leu Met Thr Gly Ser Arg Ser Thr Lys Ala Thr Pro Glu Met Asp Ser 20 25 30 Gly Leu Thr Gly Ala Thr Leu Ser Pro Lys Thr Ser Thr Gly Ala Ile 35 40 45 Val Val Thr Glu His Thr Leu Pro Phe Thr Ser Pro Asp Lys Thr Leu 50 55 60 Ala Ser Pro Thr Ser Ser Val Val Gly Arg Thr Thr Gln Ser Leu Gly 65 70 75 80 Val Met Ser Ser Ala Leu Pro Glu Ser Thr Ser Arg Gly Met Thr His 85 90 95 Ser Glu Gln Arg Thr Ser Pro Ser Leu Ser Pro Gln Val Asn Gly Thr 100 105 110 Pro Ser Arg Asn Tyr Pro Ala Thr Ser Met Val Ser Gly Leu Ser Ser 115 120 125 Pro Arg Thr Arg Thr Ser Ser Thr Glu Gly Asn Phe Thr Lys Glu Ala 130 135 140 Ser Thr Tyr Thr Leu Thr Val Glu Thr Thr Ser Gly Pro Val Thr Glu 145 150 155 160 Lys Tyr Thr Val Pro Thr Glu Thr Ser Thr Thr Glu Gly Asp Ser Thr 165 170 175 Glu Thr Pro Trp Asp Thr Arg Tyr Ile Pro Val Lys Ile Thr Ser Pro 180 185 190 Met Lys Thr Phe Ala Asp Ser Thr Ala Ser Lys Glu Asn Ala Pro Val 195 200 205 Ser Met Thr Pro Ala Glu Thr Thr Val Thr Asp Ser His Thr Pro Gly 210 215 220 Arg Thr Asn Pro Ser Phe Gly Thr Leu Tyr Ser Ser Phe Leu Asp Leu 225 230 235 240 Ser Pro Lys Gly Thr Pro Asn Ser Arg Gly Glu Thr Ser Leu Glu Leu 245 250 255 Ile Leu Ser Thr Thr Gly Tyr Pro Phe Ser Ser Pro Glu Pro Gly Ser 260 265 270 Ala Gly His Ser Arg Ile Ser Thr Ser Ala Pro Leu Ser Ser Ser Ala 275 280 285 Ser Val Leu Asp Asn Lys Ile Ser Glu Thr Ser Ile Phe Ser Gly Gln 290 295 300 Ser Leu Thr Ser Pro Leu Ser Pro Gly Val Pro Glu Ala Arg Ala Ser 305 310 315 320 Thr Met Pro Asn Ser Ala Ile Pro Phe Ser Met Thr Leu Ser Asn Ala 325 330 335 Glu Thr Ser Ala Glu Arg Val Arg Ser Thr Ile Ser Ser Leu Gly Thr 340 345 350 Pro Ser Ile Ser Thr Lys Gln Thr Ala Glu Thr Ile Leu Thr Phe His 355 360 365 Ala Phe Ala Glu Thr Met Asp Ile Pro Ser Thr His Ile Ala Lys Thr 370 375 380 Leu Ala Ser Glu Trp Leu Gly Ser Pro Gly Thr Leu Gly Gly Thr Ser 385 390 395 400 Thr Ser Ala Leu Thr Thr Thr Ser Pro Ser Thr Thr Leu Val Ser Glu 405 410 415 Glu Thr Asn Thr His His Ser Thr Ser Gly Lys Glu Thr Glu Gly Thr 420 425 430 Leu Asn Thr Ser Met Thr Pro Leu Glu Thr Ser Ala Pro Gly Glu Glu 435 440 445 Ser Glu Met Thr Ala Thr Leu Val Pro Thr Leu Gly Phe Thr Thr Leu 450 455 460 Asp Ser Lys Ile Arg Ser Pro Ser Gln Val Ser Ser Ser His Pro Thr 465 470 475 480 Arg Glu Leu Arg Thr Thr Gly Ser Thr Ser Gly Arg Gln Ser Ser Ser 485 490 495 Thr Ala Ala His Gly Ser Ser Asp Ile Leu Arg Ala Thr Thr Ser Ser 500 505 510 Thr Ser Lys Ala Ser Ser Trp Thr Ser Glu Ser Thr Ala Gln Gln Phe 515 520 525 Ser Glu Pro Gln His Thr Gln Trp Val Glu Thr Ser Pro Ser Met Lys 530 535 540 Thr Glu Arg Pro Pro Ala Ser Thr Ser Val Ala Ala Pro Ile Thr Thr 545 550 555 560 Ser Val Pro Ser Val Val Ser Gly Phe Thr Thr Leu Lys Thr Ser Ser 565 570 575 Thr Lys Gly Ile Trp Leu Glu Glu Thr Ser Ala Asp Thr Leu Ile Gly 580 585 590 Glu Ser Thr Ala Gly Pro Thr Thr His Gln Phe Ala Val Pro Thr Gly 595 600 605 Ile Ser Met Thr Gly Gly Ser Ser Thr Arg Gly Ser Gln Gly Thr Thr 610 615 620 His Leu Leu Thr Arg Ala Thr Ala Ser Ser Glu Thr Ser Ala Asp Leu 625 630 635 640 Thr Leu Ala Thr Asn Gly Val Pro Val Ser Val Ser Pro Ala Val Ser 645 650 655 Lys Thr Ala Ala Gly Ser Ser Pro Pro Gly Gly Thr Lys Pro Ser Tyr 660 665 670 Thr Met Val Ser Ser Val Ile Pro Glu Thr Ser Ser Leu Gln Ser Ser 675 680 685 Ala Phe Arg Glu Gly Thr Ser Leu Gly Leu Thr Pro Leu Asn Thr Arg 690 695 700 His Pro Phe Ser Ser Pro Glu Pro Asp Ser Ala Gly His Thr Lys Ile 705 710 715 720 Ser Thr Ser Ile Pro Leu Leu Ser Ser Ala Ser Val Leu Glu Asp Lys 725 730 735 Val Ser Ala Thr Ser Thr Phe Ser His His Lys Ala Thr Ser Ser Ile 740 745 750 Thr Thr Gly Thr Pro Glu Ile Ser Thr Lys Thr Lys Pro Ser Ser Ala 755 760 765 Val Leu Ser Ser Met Thr Leu Ser Asn Ala Ala Thr Ser Pro Glu Arg 770 775 780 Val Arg Asn Ala Thr Ser Pro Leu Thr His Pro Ser Pro Ser Gly Glu 785 790 795 800 Glu Thr Ala Gly Ser Val Leu Thr Leu Ser Thr Ser Ala Glu Thr Thr 805 810 815 Asp Ser Pro Asn Ile His Pro Thr Gly Thr Leu Thr Ser Glu Ser Ser 820 825 830 Glu Ser Pro Ser Thr Leu Ser Leu Pro Ser Val Ser Gly Val Lys Thr 835 840 845 Thr Phe Ser Ser Ser Thr Pro Ser Thr His Leu Phe Thr Ser Gly Glu 850 855 860 Glu Thr Glu Glu Thr Ser Asn Pro Ser Val Ser Gln Pro Glu Thr Ser 865 870 875 880 Val Ser Arg Val Arg Thr Thr Leu Ala Ser Thr Ser Val Pro Thr Pro 885 890 895 Val Phe Pro Thr Met Asp Thr Trp Pro Thr Arg Ser Ala Gln Phe Ser 900 905 910 Ser Ser His Leu Val Ser Glu Leu Arg Ala Thr Ser Ser Thr Ser Val 915 920 925 Thr Asn Ser Thr Gly Ser Ala Leu Pro Lys Ile Ser His Leu Thr Gly 930 935 940 Thr Ala Thr Met Ser Gln Thr Asn Arg Asp Thr Phe Asn Asp Ser Ala 945 950 955 960 Ala Pro Gln Ser Thr Thr Trp Pro Glu Thr Ser Pro Arg Phe Lys Thr 965 970 975 Gly Leu Pro Ser Ala Thr Thr Thr Val Ser Thr Ser Ala Thr Ser Leu 980 985 990 Ser Ala Thr Val Met Val Ser Lys Phe Thr Ser Pro Ala Thr Ser Ser 995 1000 1005 Met Glu Ala Thr Ser Ile Arg Glu Pro Ser Thr Thr Ile Leu Thr 1010 1015 1020 Thr Glu Thr Thr Asn Gly Pro Gly Ser Met Ala Val Ala Ser Thr 1025 1030 1035 Asn Ile Pro Ile Gly Lys Gly Tyr Ile Thr Glu Gly Arg Leu Asp 1040 1045 1050 Thr Ser His Leu Pro Ile Gly Thr Thr Ala Ser Ser Glu Thr Ser 1055 1060 1065 Met Asp Phe Thr Met Ala Lys Glu Ser Val Ser Met Ser Val Ser 1070 1075 1080 Pro Ser Gln Ser Met Asp Ala Ala Gly Ser Ser Thr Pro Gly Arg 1085 1090 1095 Thr Ser Gln Phe Val Asp Thr Phe Ser Asp Asp Val Tyr His Leu 1100 1105 1110 Thr Ser Arg Glu Ile Thr Ile Pro Arg Asp Gly Thr Ser Ser Ala 1115 1120 1125 Leu Thr Pro Gln Met Thr Ala Thr His Pro Pro Ser Pro Asp Pro 1130 1135 1140 Gly Ser Ala Arg Ser Thr Trp Leu Gly Ile Leu Ser Ser Ser Pro 1145 1150 1155 Ser Ser Pro Thr Pro Lys Val Thr Met Ser Ser Thr Phe Ser Thr 1160 1165 1170 Gln Arg Val Thr Thr Ser Met Ile Met Asp Thr Val Glu Thr Ser 1175 1180 1185 Arg Trp Asn Met Pro Asn Leu Pro Ser Thr Thr Ser Leu Thr Pro 1190 1195 1200 Ser Asn Ile Pro Thr Ser Gly Ala Ile Gly Lys Ser Thr Leu Val 1205 1210 1215 Pro Leu Asp Thr Pro Ser Pro Ala Thr Ser Leu Glu Ala Ser Glu 1220 1225 1230 Gly Gly Leu Pro Thr Leu Ser Thr Tyr Pro Glu Ser Thr Asn Thr 1235 1240 1245 Pro Ser Ile His Leu Gly Ala His Ala Ser Ser Glu Ser Pro Ser 1250 1255 1260 Thr Ile Lys Leu Thr Met Ala Ser Val Val Lys Pro Gly Ser Tyr 1265 1270 1275 Thr Pro Leu Thr Phe Pro Ser Ile Glu Thr His Ile His Val Ser 1280 1285 1290 Thr Ala Arg Met Ala Tyr Ser Ser Gly Ser Ser Pro Glu Met Thr 1295 1300 1305 Ala Pro Gly Glu Thr Asn Thr Gly Ser Thr Trp Asp Pro Thr Thr 1310 1315 1320 Tyr Ile Thr Thr Thr Asp Pro Lys Asp Thr Ser Ser Ala Gln Val 1325 1330 1335 Ser Thr Pro His Ser Val Arg Thr Leu Arg Thr Thr Glu Asn His 1340 1345 1350 Pro Lys Thr Glu Ser Ala Thr Pro Ala Ala Tyr Ser Gly Ser Pro 1355 1360 1365 Lys Ile Ser Ser Ser Pro Asn Leu Thr Ser Pro Ala Thr Lys Ala 1370 1375 1380 Trp Thr Ile Thr Asp Thr Thr Glu His Ser Thr Gln Leu His Tyr 1385 1390 1395 Thr Lys Leu Ala Glu Lys Ser Ser Gly Phe Glu Thr Gln Ser Ala 1400 1405 1410 Pro Gly Pro Val Ser Val Val Ile Pro Thr Ser Pro Thr Ile Gly 1415 1420 1425 Ser Ser Thr Leu Glu Leu Thr Ser Asp Val Pro Gly Glu Pro Leu 1430 1435 1440 Val Leu Ala Pro Ser Glu Gln Thr Thr Ile Thr Leu Pro Met Ala 1445 1450 1455 Thr Trp Leu Ser Thr Ser Leu Thr Glu Glu Met Ala Ser Thr Asp 1460 1465 1470 Leu Asp Ile Ser Ser Pro Ser Ser Pro Met Ser Thr Phe Ala Ile 1475 1480 1485 Phe Pro Pro Met Ser Thr Pro Ser His Glu Leu Ser Lys Ser Glu 1490 1495 1500 Ala Asp Thr Ser Ala Ile Arg Asn Thr Asp Ser Thr Thr Leu Asp 1505 1510 1515 Gln His Leu Gly Ile Arg Ser Leu Gly Arg Thr Gly Asp Leu Thr 1520 1525 1530 Thr Val Pro Ile Thr Pro Leu Thr Thr Thr Trp Thr Ser Val Ile 1535 1540 1545 Glu His Ser Thr Gln Ala Gln Asp Thr Leu Ser Ala Thr Met Ser 1550 1555 1560 Pro Thr His Val Thr Gln Ser Leu Lys Asp Gln Thr Ser Ile Pro 1565 1570 1575 Ala Ser Ala Ser Pro Ser His Leu Thr Glu Val Tyr Pro Glu Leu 1580 1585 1590 Gly Thr Gln Gly Arg Ser Ser Ser Glu Ala Thr Thr Phe Trp Lys 1595 1600 1605 Pro Ser Thr Asp Thr Leu Ser Arg Glu Ile Glu Thr Gly Pro Thr 1610 1615 1620 Asn Ile Gln Ser Thr Pro Pro Met Asp Asn Thr Thr Thr Gly Ser 1625 1630 1635 Ser Ser Ser Gly Val Thr Leu Gly Ile Ala His Leu Pro Ile Gly 1640 1645 1650 Thr Ser Ser Pro Ala Glu Thr Ser Thr Asn Met Ala Leu Glu Arg 1655 1660 1665 Arg Ser Ser Thr Ala Thr Val Ser Met Ala Gly Thr Met Gly Leu 1670 1675 1680 Leu Val Thr Ser Ala Pro Gly Arg Ser Ile Ser Gln Ser Leu Gly 1685 1690 1695 Arg Val Ser Ser Val Leu Ser Glu Ser Thr Thr Glu Gly Val Thr 1700 1705 1710 Asp Ser Ser Lys Gly Ser Ser Pro Arg Leu Asn Thr Gln Gly Asn 1715 1720 1725 Thr Ala Leu Ser Ser Ser Leu Glu Pro Ser Tyr Ala Glu Gly Ser 1730 1735 1740 Gln Met Ser Thr Ser Ile Pro Leu Thr Ser Ser Pro Thr Thr Pro 1745 1750 1755 Asp Val Glu Phe Ile Gly Gly Ser Thr Phe Trp Thr Lys Glu Val 1760 1765 1770 Thr Thr Val Met Thr Ser Asp Ile Ser Lys Ser Ser Ala Arg Thr 1775 1780 1785 Glu Ser Ser Ser Ala Thr Leu Met Ser Thr Ala Leu Gly Ser Thr 1790 1795 1800 Glu Asn Thr Gly Lys Glu Lys Leu Arg Thr Ala Ser Met Asp Leu 1805 1810 1815 Pro Ser Pro Thr Pro Ser Met Glu Val Thr Pro Trp Ile Ser Leu 1820 1825 1830 Thr Leu Ser Asn Ala Pro Asn Thr Thr Asp Ser Leu Asp Leu Ser 1835 1840 1845 His Gly Val His Thr Ser Ser Ala Gly Thr Leu Ala Thr Asp Arg 1850 1855 1860 Ser Leu Asn Thr Gly Val Thr Arg Ala Ser

Arg Leu Glu Asn Gly 1865 1870 1875 Ser Asp Thr Ser Ser Lys Ser Leu Ser Met Gly Asn Ser Thr His 1880 1885 1890 Thr Ser Met Thr Tyr Thr Glu Lys Ser Glu Val Ser Ser Ser Ile 1895 1900 1905 His Pro Arg Pro Glu Thr Ser Ala Pro Gly Ala Glu Thr Thr Leu 1910 1915 1920 Thr Ser Thr Pro Gly Asn Arg Ala Ile Ser Leu Thr Leu Pro Phe 1925 1930 1935 Ser Ser Ile Pro Val Glu Glu Val Ile Ser Thr Gly Ile Thr Ser 1940 1945 1950 Gly Pro Asp Ile Asn Ser Ala Pro Met Thr His Ser Pro Ile Thr 1955 1960 1965 Pro Pro Thr Ile Val Trp Thr Ser Thr Gly Thr Ile Glu Gln Ser 1970 1975 1980 Thr Gln Pro Leu His Ala Val Ser Ser Glu Lys Val Ser Val Gln 1985 1990 1995 Thr Gln Ser Thr Pro Tyr Val Asn Ser Val Ala Val Ser Ala Ser 2000 2005 2010 Pro Thr His Glu Asn Ser Val Ser Ser Gly Ser Ser Thr Ser Ser 2015 2020 2025 Pro Tyr Ser Ser Ala Ser Leu Glu Ser Leu Asp Ser Thr Ile Ser 2030 2035 2040 Arg Arg Asn Ala Ile Thr Ser Trp Leu Trp Asp Leu Thr Thr Ser 2045 2050 2055 Leu Pro Thr Thr Thr Trp Pro Ser Thr Ser Leu Ser Glu Ala Leu 2060 2065 2070 Ser Ser Gly His Ser Gly Val Ser Asn Pro Ser Ser Thr Thr Thr 2075 2080 2085 Glu Phe Pro Leu Phe Ser Ala Ala Ser Thr Ser Ala Ala Lys Gln 2090 2095 2100 Arg Asn Pro Glu Thr Glu Thr His Gly Pro Gln Asn Thr Ala Ala 2105 2110 2115 Ser Thr Leu Asn Thr Asp Ala Ser Ser Val Thr Gly Leu Ser Glu 2120 2125 2130 Thr Pro Val Gly Ala Ser Ile Ser Ser Glu Val Pro Leu Pro Met 2135 2140 2145 Ala Ile Thr Ser Arg Ser Asp Val Ser Gly Leu Thr Ser Glu Ser 2150 2155 2160 Thr Ala Asn Pro Ser Leu Gly Thr Ala Ser Ser Ala Gly Thr Lys 2165 2170 2175 Leu Thr Arg Thr Ile Ser Leu Pro Thr Ser Glu Ser Leu Val Ser 2180 2185 2190 Phe Arg Met Asn Lys Asp Pro Trp Thr Val Ser Ile Pro Leu Gly 2195 2200 2205 Ser His Pro Thr Thr Asn Thr Glu Thr Ser Ile Pro Val Asn Ser 2210 2215 2220 Ala Gly Pro Pro Gly Leu Ser Thr Val Ala Ser Asp Val Ile Asp 2225 2230 2235 Thr Pro Ser Asp Gly Ala Glu Ser Ile Pro Thr Val Ser Phe Ser 2240 2245 2250 Pro Ser Pro Asp Thr Glu Val Thr Thr Ile Ser His Phe Pro Glu 2255 2260 2265 Lys Thr Thr His Ser Phe Arg Thr Ile Ser Ser Leu Thr His Glu 2270 2275 2280 Leu Thr Ser Arg Val Thr Pro Ile Pro Gly Asp Trp Met Ser Ser 2285 2290 2295 Ala Met Ser Thr Lys Pro Thr Gly Ala Ser Pro Ser Ile Thr Leu 2300 2305 2310 Gly Glu Arg Arg Thr Ile Thr Ser Ala Ala Pro Thr Thr Ser Pro 2315 2320 2325 Ile Val Leu Thr Ala Ser Phe Thr Glu Thr Ser Thr Val Ser Leu 2330 2335 2340 Asp Asn Glu Thr Thr Val Lys Thr Ser Asp Ile Leu Asp Ala Arg 2345 2350 2355 Lys Thr Asn Glu Leu Pro Ser Asp Ser Ser Ser Ser Ser Asp Leu 2360 2365 2370 Ile Asn Thr Ser Ile Ala Ser Ser Thr Met Asp Val Thr Lys Thr 2375 2380 2385 Ala Ser Ile Ser Pro Thr Ser Ile Ser Gly Met Thr Ala Ser Ser 2390 2395 2400 Ser Pro Ser Leu Phe Ser Ser Asp Arg Pro Gln Val Pro Thr Ser 2405 2410 2415 Thr Thr Glu Thr Asn Thr Ala Thr Ser Pro Ser Val Ser Ser Asn 2420 2425 2430 Thr Tyr Ser Leu Asp Gly Gly Ser Asn Val Gly Gly Thr Pro Ser 2435 2440 2445 Thr Leu Pro Pro Phe Thr Ile Thr His Pro Val Glu Thr Ser Ser 2450 2455 2460 Ala Leu Leu Ala Trp Ser Arg Pro Val Arg Thr Phe Ser Thr Met 2465 2470 2475 Val Ser Thr Asp Thr Ala Ser Gly Glu Asn Pro Thr Ser Ser Asn 2480 2485 2490 Ser Val Val Thr Ser Val Pro Ala Pro Gly Thr Trp Thr Ser Val 2495 2500 2505 Gly Ser Thr Thr Asp Leu Pro Ala Met Gly Phe Leu Lys Thr Ser 2510 2515 2520 Pro Ala Gly Glu Ala His Ser Leu Leu Ala Ser Thr Ile Glu Pro 2525 2530 2535 Ala Thr Ala Phe Thr Pro His Leu Ser Ala Ala Val Val Thr Gly 2540 2545 2550 Ser Ser Ala Thr Ser Glu Ala Ser Leu Leu Thr Thr Ser Glu Ser 2555 2560 2565 Lys Ala Ile His Ser Ser Pro Gln Thr Pro Thr Thr Pro Thr Ser 2570 2575 2580 Gly Ala Asn Trp Glu Thr Ser Ala Thr Pro Glu Ser Leu Leu Val 2585 2590 2595 Val Thr Glu Thr Ser Asp Thr Thr Leu Thr Ser Lys Ile Leu Val 2600 2605 2610 Thr Asp Thr Ile Leu Phe Ser Thr Val Ser Thr Pro Pro Ser Lys 2615 2620 2625 Phe Pro Ser Thr Gly Thr Leu Ser Gly Ala Ser Phe Pro Thr Leu 2630 2635 2640 Leu Pro Asp Thr Pro Ala Ile Pro Leu Thr Ala Thr Glu Pro Thr 2645 2650 2655 Ser Ser Leu Ala Thr Ser Phe Asp Ser Thr Pro Leu Val Thr Ile 2660 2665 2670 Ala Ser Asp Ser Leu Gly Thr Val Pro Glu Thr Thr Leu Thr Met 2675 2680 2685 Ser Glu Thr Ser Asn Gly Asp Ala Leu Val Leu Lys Thr Val Ser 2690 2695 2700 Asn Pro Asp Arg Ser Ile Pro Gly Ile Thr Ile Gln Gly Val Thr 2705 2710 2715 Glu Ser Pro Leu His Pro Ser Ser Thr Ser Pro Ser Lys Ile Val 2720 2725 2730 Ala Pro Arg Asn Thr Thr Tyr Glu Gly Ser Ile Thr Val Ala Leu 2735 2740 2745 Ser Thr Leu Pro Ala Gly Thr Thr Gly Ser Leu Val Phe Ser Gln 2750 2755 2760 Ser Ser Glu Asn Ser Glu Thr Thr Ala Leu Val Asp Ser Ser Ala 2765 2770 2775 Gly Leu Glu Arg Ala Ser Val Met Pro Leu Thr Thr Gly Ser Gln 2780 2785 2790 Gly Met Ala Ser Ser Gly Gly Ile Arg Ser Gly Ser Thr His Ser 2795 2800 2805 Thr Gly Thr Lys Thr Phe Ser Ser Leu Pro Leu Thr Met Asn Pro 2810 2815 2820 Gly Glu Val Thr Ala Met Ser Glu Ile Thr Thr Asn Arg Leu Thr 2825 2830 2835 Ala Thr Gln Ser Thr Ala Pro Lys Gly Ile Pro Val Lys Pro Thr 2840 2845 2850 Ser Ala Glu Ser Gly Leu Leu Thr Pro Val Ser Ala Ser Ser Ser 2855 2860 2865 Pro Ser Lys Ala Phe Ala Ser Leu Thr Thr Ala Pro Pro Thr Trp 2870 2875 2880 Gly Ile Pro Gln Ser Thr Leu Thr Phe Glu Phe Ser Glu Val Pro 2885 2890 2895 Ser Leu Asp Thr Lys Ser Ala Ser Leu Pro Thr Pro Gly Gln Ser 2900 2905 2910 Leu Asn Thr Ile Pro Asp Ser Asp Ala Ser Thr Ala Ser Ser Ser 2915 2920 2925 Leu Ser Lys Ser Pro Glu Lys Asn Pro Arg Ala Arg Met Met Thr 2930 2935 2940 Ser Thr Lys Ala Ile Ser Ala Ser Ser Phe Gln Ser Thr Gly Phe 2945 2950 2955 Thr Glu Thr Pro Glu Gly Ser Ala Ser Pro Ser Met Ala Gly His 2960 2965 2970 Glu Pro Arg Val Pro Thr Ser Gly Thr Gly Asp Pro Arg Tyr Ala 2975 2980 2985 Ser Glu Ser Met Ser Tyr Pro Asp Pro Ser Lys Ala Ser Ser Ala 2990 2995 3000 Met Thr Ser Thr Ser Leu Ala Ser Lys Leu Thr Thr Leu Phe Ser 3005 3010 3015 Thr Gly Gln Ala Ala Arg Ser Gly Ser Ser Ser Ser Pro Ile Ser 3020 3025 3030 Leu Ser Thr Glu Lys Glu Thr Ser Phe Leu Ser Pro Thr Ala Ser 3035 3040 3045 Thr Ser Arg Lys Thr Ser Leu Phe Leu Gly Pro Ser Met Ala Arg 3050 3055 3060 Gln Pro Asn Ile Leu Val His Leu Gln Thr Ser Ala Leu Thr Leu 3065 3070 3075 Ser Pro Thr Ser Thr Leu Asn Met Ser Gln Glu Glu Pro Pro Glu 3080 3085 3090 Leu Thr Ser Ser Gln Thr Ile Ala Glu Glu Glu Gly Thr Thr Ala 3095 3100 3105 Glu Thr Gln Thr Leu Thr Phe Thr Pro Ser Glu Thr Pro Thr Ser 3110 3115 3120 Leu Leu Pro Val Ser Ser Pro Thr Glu Pro Thr Ala Arg Arg Lys 3125 3130 3135 Ser Ser Pro Glu Thr Trp Ala Ser Ser Ile Ser Val Pro Ala Lys 3140 3145 3150 Thr Ser Leu Val Glu Thr Thr Asp Gly Thr Leu Val Thr Thr Ile 3155 3160 3165 Lys Met Ser Ser Gln Ala Ala Gln Gly Asn Ser Thr Trp Pro Ala 3170 3175 3180 Pro Ala Glu Glu Thr Gly Ser Ser Pro Ala Gly Thr Ser Pro Gly 3185 3190 3195 Ser Pro Glu Met Ser Thr Thr Leu Lys Ile Met Ser Ser Lys Glu 3200 3205 3210 Pro Ser Ile Ser Pro Glu Ile Arg Ser Thr Val Arg Asn Ser Pro 3215 3220 3225 Trp Lys Thr Pro Glu Thr Thr Val Pro Met Glu Thr Thr Val Glu 3230 3235 3240 Pro Val Thr Leu Gln Ser Thr Ala Leu Gly Ser Gly Ser Thr Ser 3245 3250 3255 Ile Ser His Leu Pro Thr Gly Thr Thr Ser Pro Thr Lys Ser Pro 3260 3265 3270 Thr Glu Asn Met Leu Ala Thr Glu Arg Val Ser Leu Ser Pro Ser 3275 3280 3285 Pro Pro Glu Ala Trp Thr Asn Leu Tyr Ser Gly Thr Pro Gly Gly 3290 3295 3300 Thr Arg Gln Ser Leu Ala Thr Met Ser Ser Val Ser Leu Glu Ser 3305 3310 3315 Pro Thr Ala Arg Ser Ile Thr Gly Thr Gly Gln Gln Ser Ser Pro 3320 3325 3330 Glu Leu Val Ser Lys Thr Thr Gly Met Glu Phe Ser Met Trp His 3335 3340 3345 Gly Ser Thr Gly Gly Thr Thr Gly Asp Thr His Val Ser Leu Ser 3350 3355 3360 Thr Ser Ser Asn Ile Leu Glu Asp Pro Val Thr Ser Pro Asn Ser 3365 3370 3375 Val Ser Ser Leu Thr Asp Lys Ser Lys His Lys Thr Glu Thr Trp 3380 3385 3390 Val Ser Thr Thr Ala Ile Pro Ser Thr Val Leu Asn Asn Lys Ile 3395 3400 3405 Met Ala Ala Glu Gln Gln Thr Ser Arg Ser Val Asp Glu Ala Tyr 3410 3415 3420 Ser Ser Thr Ser Ser Trp Ser Asp Gln Thr Ser Gly Ser Asp Ile 3425 3430 3435 Thr Leu Gly Ala Ser Pro Asp Val Thr Asn Thr Leu Tyr Ile Thr 3440 3445 3450 Ser Thr Ala Gln Thr Thr Ser Leu Val Ser Leu Pro Ser Gly Asp 3455 3460 3465 Gln Gly Ile Thr Ser Leu Thr Asn Pro Ser Gly Gly Lys Thr Ser 3470 3475 3480 Ser Ala Ser Ser Val Thr Ser Pro Ser Ile Gly Leu Glu Thr Leu 3485 3490 3495 Arg Ala Asn Val Ser Ala Val Lys Ser Asp Ile Ala Pro Thr Ala 3500 3505 3510 Gly His Leu Ser Gln Thr Ser Ser Pro Ala Glu Val Ser Ile Leu 3515 3520 3525 Asp Val Thr Thr Ala Pro Thr Pro Gly Ile Ser Thr Thr Ile Thr 3530 3535 3540 Thr Met Gly Thr Asn Ser Ile Ser Thr Thr Thr Pro Asn Pro Glu 3545 3550 3555 Val Gly Met Ser Thr Met Asp Ser Thr Pro Ala Thr Glu Arg Arg 3560 3565 3570 Thr Thr Ser Thr Glu His Pro Ser Thr Trp Ser Ser Thr Ala Ala 3575 3580 3585 Ser Asp Ser Trp Thr Val Thr Asp Met Thr Ser Asn Leu Lys Val 3590 3595 3600 Ala Arg Ser Pro Gly Thr Ile Ser Thr Met His Thr Thr Ser Phe 3605 3610 3615 Leu Ala Ser Ser Thr Glu Leu Asp Ser Met Ser Thr Pro His Gly 3620 3625 3630 Arg Ile Thr Val Ile Gly Thr Ser Leu Val Thr Pro Ser Ser Asp 3635 3640 3645 Ala Ser Ala Val Lys Thr Glu Thr Ser Thr Ser Glu Arg Thr Leu 3650 3655 3660 Ser Pro Ser Asp Thr Thr Ala Ser Thr Pro Ile Ser Thr Phe Ser 3665 3670 3675 Arg Val Gln Arg Met Ser Ile Ser Val Pro Asp Ile Leu Ser Thr 3680 3685 3690 Ser Trp Thr Pro Ser Ser Thr Glu Ala Glu Asp Val Pro Val Ser 3695 3700 3705 Met Val Ser Thr Asp His Ala Ser Thr Lys Thr Asp Pro Asn Thr 3710 3715 3720 Pro Leu Ser Thr Phe Leu Phe Asp Ser Leu Ser Thr Leu Asp Trp 3725 3730 3735 Asp Thr Gly Arg Ser Leu Ser Ser Ala Thr Ala Thr Thr Ser Ala 3740 3745 3750 Pro Gln Gly Ala Thr Thr Pro Gln Glu Leu Thr Leu Glu Thr Met 3755 3760 3765 Ile Ser Pro Ala Thr Ser Gln Leu Pro Phe Ser Ile Gly His Ile 3770 3775 3780 Thr Ser Ala Val Thr Pro Ala Ala Met Ala Arg Ser Ser Gly Val 3785 3790 3795 Thr Phe Ser Arg Pro Asp Pro Thr Ser Lys Lys Ala Glu Gln Thr 3800 3805 3810 Ser Thr Gln Leu Pro Thr Thr Thr Ser Ala His Pro Gly Gln Val 3815 3820 3825 Pro Arg Ser Ala Ala Thr Thr Leu Asp Val Ile Pro His Thr Ala 3830 3835 3840 Lys Thr Pro Asp Ala Thr Phe Gln Arg Gln Gly Gln Thr Ala Leu 3845 3850 3855 Thr Thr Glu Ala Arg Ala Thr Ser Asp Ser Trp Asn Glu Lys Glu 3860 3865 3870 Lys Ser Thr Pro Ser Ala Pro Trp Ile Thr Glu Met Met Asn Ser 3875 3880 3885 Val Ser Glu Asp Thr Ile Lys Glu Val Thr Ser Ser Ser Ser Val 3890 3895 3900 Leu Arg Thr Leu Asn Thr Leu Asp Ile Asn Leu Glu Ser Gly Thr 3905 3910 3915 Thr Ser Ser Pro Ser Trp Lys Ser Ser Pro Tyr Glu Arg Ile Ala 3920 3925 3930 Pro Ser Glu Ser Thr Thr Asp Lys Glu Ala Ile His Pro Ser Thr 3935 3940 3945 Asn Thr Val Glu Thr Thr Gly Trp Val Thr Ser Ser Glu His Ala 3950 3955 3960 Ser His Ser Thr Ile Pro Ala His Ser Ala Ser Ser Lys Leu Thr 3965 3970 3975 Ser Pro Val Val Thr Thr Ser Thr Arg Glu Gln Ala Ile Val Ser 3980 3985 3990 Met Ser Thr Thr Thr Trp Pro Glu Ser Thr Arg Ala Arg Thr Glu 3995 4000 4005 Pro Asn Ser Phe Leu Thr Ile Glu Leu Arg Asp Val Ser Pro Tyr 4010 4015 4020 Met Asp Thr Ser Ser Thr Thr Gln Thr Ser Ile Ile Ser Ser Pro 4025 4030 4035 Gly Ser Thr Ala Ile Thr Lys Gly Pro Arg Thr Glu Ile Thr Ser 4040 4045 4050 Ser Lys Arg Ile Ser Ser Ser Phe Leu Ala Gln Ser Met Arg Ser 4055 4060

4065 Ser Asp Ser Pro Ser Glu Ala Ile Thr Arg Leu Ser Asn Phe Pro 4070 4075 4080 Ala Met Thr Glu Ser Gly Gly Met Ile Leu Ala Met Gln Thr Ser 4085 4090 4095 Pro Pro Gly Ala Thr Ser Leu Ser Ala Pro Thr Leu Asp Thr Ser 4100 4105 4110 Ala Thr Ala Ser Trp Thr Gly Thr Pro Leu Ala Thr Thr Gln Arg 4115 4120 4125 Phe Thr Tyr Ser Glu Lys Thr Thr Leu Phe Ser Lys Gly Pro Glu 4130 4135 4140 Asp Thr Ser Gln Pro Ser Pro Pro Ser Val Glu Glu Thr Ser Ser 4145 4150 4155 Ser Ser Ser Leu Val Pro Ile His Ala Thr Thr Ser Pro Ser Asn 4160 4165 4170 Ile Leu Leu Thr Ser Gln Gly His Ser Pro Ser Ser Thr Pro Pro 4175 4180 4185 Val Thr Ser Val Phe Leu Ser Glu Thr Ser Gly Leu Gly Lys Thr 4190 4195 4200 Thr Asp Met Ser Arg Ile Ser Leu Glu Pro Gly Thr Ser Leu Pro 4205 4210 4215 Pro Asn Leu Ser Ser Thr Ala Gly Glu Ala Leu Ser Thr Tyr Glu 4220 4225 4230 Ala Ser Arg Asp Thr Lys Ala Ile His His Ser Ala Asp Thr Ala 4235 4240 4245 Val Thr Asn Met Glu Ala Thr Ser Ser Glu Tyr Ser Pro Ile Pro 4250 4255 4260 Gly His Thr Lys Pro Ser Lys Ala Thr Ser Pro Leu Val Thr Ser 4265 4270 4275 His Ile Met Gly Asp Ile Thr Ser Ser Thr Ser Val Phe Gly Ser 4280 4285 4290 Ser Glu Thr Thr Glu Ile Glu Thr Val Ser Ser Val Asn Gln Gly 4295 4300 4305 Leu Gln Glu Arg Ser Thr Ser Gln Val Ala Ser Ser Ala Thr Glu 4310 4315 4320 Thr Ser Thr Val Ile Thr His Val Ser Ser Gly Asp Ala Thr Thr 4325 4330 4335 His Val Thr Lys Thr Gln Ala Thr Phe Ser Ser Gly Thr Ser Ile 4340 4345 4350 Ser Ser Pro His Gln Phe Ile Thr Ser Thr Asn Thr Phe Thr Asp 4355 4360 4365 Val Ser Thr Asn Pro Ser Thr Ser Leu Ile Met Thr Glu Ser Ser 4370 4375 4380 Gly Val Thr Ile Thr Thr Gln Thr Gly Pro Thr Gly Ala Ala Thr 4385 4390 4395 Gln Gly Pro Tyr Leu Leu Asp Thr Ser Thr Met Pro Tyr Leu Thr 4400 4405 4410 Glu Thr Pro Leu Ala Val Thr Pro Asp Phe Met Gln Ser Glu Lys 4415 4420 4425 Thr Thr Leu Ile Ser Lys Gly Pro Lys Asp Val Ser Trp Thr Ser 4430 4435 4440 Pro Pro Ser Val Ala Glu Thr Ser Tyr Pro Ser Ser Leu Thr Pro 4445 4450 4455 Phe Leu Val Thr Thr Ile Pro Pro Ala Thr Ser Thr Leu Gln Gly 4460 4465 4470 Gln His Thr Ser Ser Pro Val Ser Ala Thr Ser Val Leu Thr Ser 4475 4480 4485 Gly Leu Val Lys Thr Thr Asp Met Leu Asn Thr Ser Met Glu Pro 4490 4495 4500 Val Thr Asn Ser Pro Gln Asn Leu Asn Asn Pro Ser Asn Glu Ile 4505 4510 4515 Leu Ala Thr Leu Ala Ala Thr Thr Asp Ile Glu Thr Ile His Pro 4520 4525 4530 Ser Ile Asn Lys Ala Val Thr Asn Met Gly Thr Ala Ser Ser Ala 4535 4540 4545 His Val Leu His Ser Thr Leu Pro Val Ser Ser Glu Pro Ser Thr 4550 4555 4560 Ala Thr Ser Pro Met Val Pro Ala Ser Ser Met Gly Asp Ala Leu 4565 4570 4575 Ala Ser Ile Ser Ile Pro Gly Ser Glu Thr Thr Asp Ile Glu Gly 4580 4585 4590 Glu Pro Thr Ser Ser Leu Thr Ala Gly Arg Lys Glu Asn Ser Thr 4595 4600 4605 Leu Gln Glu Met Asn Ser Thr Thr Glu Ser Asn Ile Ile Leu Ser 4610 4615 4620 Asn Val Ser Val Gly Ala Ile Thr Glu Ala Thr Lys Met Glu Val 4625 4630 4635 Pro Ser Phe Asp Ala Thr Phe Ile Pro Thr Pro Ala Gln Ser Thr 4640 4645 4650 Lys Phe Pro Asp Ile Phe Ser Val Ala Ser Ser Arg Leu Ser Asn 4655 4660 4665 Ser Pro Pro Met Thr Ile Ser Thr His Met Thr Thr Thr Gln Thr 4670 4675 4680 Gly Ser Ser Gly Ala Thr Ser Lys Ile Pro Leu Ala Leu Asp Thr 4685 4690 4695 Ser Thr Leu Glu Thr Ser Ala Gly Thr Pro Ser Val Val Thr Glu 4700 4705 4710 Gly Phe Ala His Ser Lys Ile Thr Thr Ala Met Asn Asn Asp Val 4715 4720 4725 Lys Asp Val Ser Gln Thr Asn Pro Pro Phe Gln Asp Glu Ala Ser 4730 4735 4740 Ser Pro Ser Ser Gln Ala Pro Val Leu Val Thr Thr Leu Pro Ser 4745 4750 4755 Ser Val Ala Phe Thr Pro Gln Trp His Ser Thr Ser Ser Pro Val 4760 4765 4770 Ser Met Ser Ser Val Leu Thr Ser Ser Leu Val Lys Thr Ala Gly 4775 4780 4785 Lys Val Asp Thr Ser Leu Glu Thr Val Thr Ser Ser Pro Gln Ser 4790 4795 4800 Met Ser Asn Thr Leu Asp Asp Ile Ser Val Thr Ser Ala Ala Thr 4805 4810 4815 Thr Asp Ile Glu Thr Thr His Pro Ser Ile Asn Thr Val Val Thr 4820 4825 4830 Asn Val Gly Thr Thr Gly Ser Ala Phe Glu Ser His Ser Thr Val 4835 4840 4845 Ser Ala Tyr Pro Glu Pro Ser Lys Val Thr Ser Pro Asn Val Thr 4850 4855 4860 Thr Ser Thr Met Glu Asp Thr Thr Ile Ser Arg Ser Ile Pro Lys 4865 4870 4875 Ser Ser Lys Thr Thr Arg Thr Glu Thr Glu Thr Thr Ser Ser Leu 4880 4885 4890 Thr Pro Lys Leu Arg Glu Thr Ser Ile Ser Gln Glu Ile Thr Ser 4895 4900 4905 Ser Thr Glu Thr Ser Thr Val Pro Tyr Lys Glu Leu Thr Gly Ala 4910 4915 4920 Thr Thr Glu Val Ser Arg Thr Asp Val Thr Ser Ser Ser Ser Thr 4925 4930 4935 Ser Phe Pro Gly Pro Asp Gln Ser Thr Val Ser Leu Asp Ile Ser 4940 4945 4950 Thr Glu Thr Asn Thr Arg Leu Ser Thr Ser Pro Ile Met Thr Glu 4955 4960 4965 Ser Ala Glu Ile Thr Ile Thr Thr Gln Thr Gly Pro His Gly Ala 4970 4975 4980 Thr Ser Gln Asp Thr Phe Thr Met Asp Pro Ser Asn Thr Thr Pro 4985 4990 4995 Gln Ala Gly Ile His Ser Ala Met Thr His Gly Phe Ser Gln Leu 5000 5005 5010 Asp Val Thr Thr Leu Met Ser Arg Ile Pro Gln Asp Val Ser Trp 5015 5020 5025 Thr Ser Pro Pro Ser Val Asp Lys Thr Ser Ser Pro Ser Ser Phe 5030 5035 5040 Leu Ser Ser Pro Ala Met Thr Thr Pro Ser Leu Ile Ser Ser Thr 5045 5050 5055 Leu Pro Glu Asp Lys Leu Ser Ser Pro Met Thr Ser Leu Leu Thr 5060 5065 5070 Ser Gly Leu Val Lys Ile Thr Asp Ile Leu Arg Thr Arg Leu Glu 5075 5080 5085 Pro Val Thr Ser Ser Leu Pro Asn Phe Ser Ser Thr Ser Asp Lys 5090 5095 5100 Ile Leu Ala Thr Ser Lys Asp Ser Lys Asp Thr Lys Glu Ile Phe 5105 5110 5115 Pro Ser Ile Asn Thr Glu Glu Thr Asn Val Lys Ala Asn Asn Ser 5120 5125 5130 Gly His Glu Ser His Ser Pro Ala Leu Ala Asp Ser Glu Thr Pro 5135 5140 5145 Lys Ala Thr Thr Gln Met Val Ile Thr Thr Thr Val Gly Asp Pro 5150 5155 5160 Ala Pro Ser Thr Ser Met Pro Val His Gly Ser Ser Glu Thr Thr 5165 5170 5175 Asn Ile Lys Arg Glu Pro Thr Tyr Phe Leu Thr Pro Arg Leu Arg 5180 5185 5190 Glu Thr Ser Thr Ser Gln Glu Ser Ser Phe Pro Thr Asp Thr Ser 5195 5200 5205 Phe Leu Leu Ser Lys Val Pro Thr Gly Thr Ile Thr Glu Val Ser 5210 5215 5220 Ser Thr Gly Val Asn Ser Ser Ser Lys Ile Ser Thr Pro Asp His 5225 5230 5235 Asp Lys Ser Thr Val Pro Pro Asp Thr Phe Thr Gly Glu Ile Pro 5240 5245 5250 Arg Val Phe Thr Ser Ser Ile Lys Thr Lys Ser Ala Glu Met Thr 5255 5260 5265 Ile Thr Thr Gln Ala Ser Pro Pro Glu Ser Ala Ser His Ser Thr 5270 5275 5280 Leu Pro Leu Asp Thr Ser Thr Thr Leu Ser Gln Gly Gly Thr His 5285 5290 5295 Ser Thr Val Thr Gln Gly Phe Pro Tyr Ser Glu Val Thr Thr Leu 5300 5305 5310 Met Gly Met Gly Pro Gly Asn Val Ser Trp Met Thr Thr Pro Pro 5315 5320 5325 Val Glu Glu Thr Ser Ser Val Ser Ser Leu Met Ser Ser Pro Ala 5330 5335 5340 Met Thr Ser Pro Ser Pro Val Ser Ser Thr Ser Pro Gln Ser Ile 5345 5350 5355 Pro Ser Ser Pro Leu Pro Val Thr Ala Leu Pro Thr Ser Val Leu 5360 5365 5370 Val Thr Thr Thr Asp Val Leu Gly Thr Thr Ser Pro Glu Ser Val 5375 5380 5385 Thr Ser Ser Pro Pro Asn Leu Ser Ser Ile Thr His Glu Arg Pro 5390 5395 5400 Ala Thr Tyr Lys Asp Thr Ala His Thr Glu Ala Ala Met His His 5405 5410 5415 Ser Thr Asn Thr Ala Val Thr Asn Val Gly Thr Ser Gly Ser Gly 5420 5425 5430 His Lys Ser Gln Ser Ser Val Leu Ala Asp Ser Glu Thr Ser Lys 5435 5440 5445 Ala Thr Pro Leu Met Ser Thr Thr Ser Thr Leu Gly Asp Thr Ser 5450 5455 5460 Val Ser Thr Ser Thr Pro Asn Ile Ser Gln Thr Asn Gln Ile Gln 5465 5470 5475 Thr Glu Pro Thr Ala Ser Leu Ser Pro Arg Leu Arg Glu Ser Ser 5480 5485 5490 Thr Ser Glu Lys Thr Ser Ser Thr Thr Glu Thr Asn Thr Ala Phe 5495 5500 5505 Ser Tyr Val Pro Thr Gly Ala Ile Thr Gln Ala Ser Arg Thr Glu 5510 5515 5520 Ile Ser Ser Ser Arg Thr Ser Ile Ser Asp Leu Asp Arg Pro Thr 5525 5530 5535 Ile Ala Pro Asp Ile Ser Thr Gly Met Ile Thr Arg Leu Phe Thr 5540 5545 5550 Ser Pro Ile Met Thr Lys Ser Ala Glu Met Thr Val Thr Thr Gln 5555 5560 5565 Thr Thr Thr Pro Gly Ala Thr Ser Gln Gly Ile Leu Pro Trp Asp 5570 5575 5580 Thr Ser Thr Thr Leu Phe Gln Gly Gly Thr His Ser Thr Val Ser 5585 5590 5595 Gln Gly Phe Pro His Ser Glu Ile Thr Thr Leu Arg Ser Arg Thr 5600 5605 5610 Pro Gly Asp Val Ser Trp Met Thr Thr Pro Pro Val Glu Glu Thr 5615 5620 5625 Ser Ser Gly Phe Ser Leu Met Ser Pro Ser Met Thr Ser Pro Ser 5630 5635 5640 Pro Val Ser Ser Thr Ser Pro Glu Ser Ile Pro Ser Ser Pro Leu 5645 5650 5655 Pro Val Thr Ala Leu Leu Thr Ser Val Leu Val Thr Thr Thr Asn 5660 5665 5670 Val Leu Gly Thr Thr Ser Pro Glu Pro Val Thr Ser Ser Pro Pro 5675 5680 5685 Asn Leu Ser Ser Pro Thr Gln Glu Arg Leu Thr Thr Tyr Lys Asp 5690 5695 5700 Thr Ala His Thr Glu Ala Met His Ala Ser Met His Thr Asn Thr 5705 5710 5715 Ala Val Ala Asn Val Gly Thr Ser Ile Ser Gly His Glu Ser Gln 5720 5725 5730 Ser Ser Val Pro Ala Asp Ser His Thr Ser Lys Ala Thr Ser Pro 5735 5740 5745 Met Gly Ile Thr Phe Ala Met Gly Asp Thr Ser Val Ser Thr Ser 5750 5755 5760 Thr Pro Ala Phe Phe Glu Thr Arg Ile Gln Thr Glu Ser Thr Ser 5765 5770 5775 Ser Leu Ile Pro Gly Leu Arg Asp Thr Arg Thr Ser Glu Glu Ile 5780 5785 5790 Asn Thr Val Thr Glu Thr Ser Thr Val Leu Ser Glu Val Pro Thr 5795 5800 5805 Thr Thr Thr Thr Glu Val Ser Arg Thr Glu Val Ile Thr Ser Ser 5810 5815 5820 Arg Thr Thr Ile Ser Gly Pro Asp His Ser Lys Met Ser Pro Tyr 5825 5830 5835 Ile Ser Thr Glu Thr Ile Thr Arg Leu Ser Thr Phe Pro Phe Val 5840 5845 5850 Thr Gly Ser Thr Glu Met Ala Ile Thr Asn Gln Thr Gly Pro Ile 5855 5860 5865 Gly Thr Ile Ser Gln Ala Thr Leu Thr Leu Asp Thr Ser Ser Thr 5870 5875 5880 Ala Ser Trp Glu Gly Thr His Ser Pro Val Thr Gln Arg Phe Pro 5885 5890 5895 His Ser Glu Glu Thr Thr Thr Met Ser Arg Ser Thr Lys Gly Val 5900 5905 5910 Ser Trp Gln Ser Pro Pro Ser Val Glu Glu Thr Ser Ser Pro Ser 5915 5920 5925 Ser Pro Val Pro Leu Pro Ala Ile Thr Ser His Ser Ser Leu Tyr 5930 5935 5940 Ser Ala Val Ser Gly Ser Ser Pro Thr Ser Ala Leu Pro Val Thr 5945 5950 5955 Ser Leu Leu Thr Ser Gly Arg Arg Lys Thr Ile Asp Met Leu Asp 5960 5965 5970 Thr His Ser Glu Leu Val Thr Ser Ser Leu Pro Ser Ala Ser Ser 5975 5980 5985 Phe Ser Gly Glu Ile Leu Thr Ser Glu Ala Ser Thr Asn Thr Glu 5990 5995 6000 Thr Ile His Phe Ser Glu Asn Thr Ala Glu Thr Asn Met Gly Thr 6005 6010 6015 Thr Asn Ser Met His Lys Leu His Ser Ser Val Ser Ile His Ser 6020 6025 6030 Gln Pro Ser Gly His Thr Pro Pro Lys Val Thr Gly Ser Met Met 6035 6040 6045 Glu Asp Ala Ile Val Ser Thr Ser Thr Pro Gly Ser Pro Glu Thr 6050 6055 6060 Lys Asn Val Asp Arg Asp Ser Thr Ser Pro Leu Thr Pro Glu Leu 6065 6070 6075 Lys Glu Asp Ser Thr Ala Leu Val Met Asn Ser Thr Thr Glu Ser 6080 6085 6090 Asn Thr Val Phe Ser Ser Val Ser Leu Asp Ala Ala Thr Glu Val 6095 6100 6105 Ser Arg Ala Glu Val Thr Tyr Tyr Asp Pro Thr Phe Met Pro Ala 6110 6115 6120 Ser Ala Gln Ser Thr Lys Ser Pro Asp Ile Ser Pro Glu Ala Ser 6125 6130 6135 Ser Ser His Ser Asn Ser Pro Pro Leu Thr Ile Ser Thr His Lys 6140 6145 6150 Thr Ile Ala Thr Gln Thr Gly Pro Ser Gly Val Thr Ser Leu Gly 6155 6160 6165 Gln Leu Thr Leu Asp Thr Ser Thr Ile Ala Thr Ser Ala Gly Thr 6170 6175 6180 Pro Ser Ala Arg Thr Gln Asp Phe Val Asp Ser Glu Thr Thr Ser 6185 6190 6195 Val Met Asn Asn Asp Leu Asn Asp Val Leu Lys Thr Ser Pro Phe 6200 6205 6210 Ser Ala Glu Glu Ala Asn Ser Leu Ser Ser Gln Ala Pro Leu Leu 6215 6220 6225 Val Thr Thr Ser Pro Ser Pro Val Thr Ser Thr Leu Gln Glu His 6230 6235 6240 Ser Thr Ser Ser Leu Val Ser Val Thr Ser Val Pro Thr Pro Thr 6245 6250 6255 Leu Ala Lys Ile Thr

Asp Met Asp Thr Asn Leu Glu Pro Val Thr 6260 6265 6270 Arg Ser Pro Gln Asn Leu Arg Asn Thr Leu Ala Thr Ser Glu Ala 6275 6280 6285 Thr Thr Asp Thr His Thr Met His Pro Ser Ile Asn Thr Ala Val 6290 6295 6300 Ala Asn Val Gly Thr Thr Ser Ser Pro Asn Glu Phe Tyr Phe Thr 6305 6310 6315 Val Ser Pro Asp Ser Asp Pro Tyr Lys Ala Thr Ser Ala Val Val 6320 6325 6330 Ile Thr Ser Thr Ser Gly Asp Ser Ile Val Ser Thr Ser Met Pro 6335 6340 6345 Arg Ser Ser Ala Met Lys Lys Ile Glu Ser Glu Thr Thr Phe Ser 6350 6355 6360 Leu Ile Phe Arg Leu Arg Glu Thr Ser Thr Ser Gln Lys Ile Gly 6365 6370 6375 Ser Ser Ser Asp Thr Ser Thr Val Phe Asp Lys Ala Phe Thr Ala 6380 6385 6390 Ala Thr Thr Glu Val Ser Arg Thr Glu Leu Thr Ser Ser Ser Arg 6395 6400 6405 Thr Ser Ile Gln Gly Thr Glu Lys Pro Thr Met Ser Pro Asp Thr 6410 6415 6420 Ser Thr Arg Ser Val Thr Met Leu Ser Thr Phe Ala Gly Leu Thr 6425 6430 6435 Lys Ser Glu Glu Arg Thr Ile Ala Thr Gln Thr Gly Pro His Arg 6440 6445 6450 Ala Thr Ser Gln Gly Thr Leu Thr Trp Asp Thr Ser Ile Thr Thr 6455 6460 6465 Ser Gln Ala Gly Thr His Ser Ala Met Thr His Gly Phe Ser Gln 6470 6475 6480 Leu Asp Leu Ser Thr Leu Thr Ser Arg Val Pro Glu Tyr Ile Ser 6485 6490 6495 Gly Thr Ser Pro Pro Ser Val Glu Lys Thr Ser Ser Ser Ser Ser 6500 6505 6510 Leu Leu Ser Leu Pro Ala Ile Thr Ser Pro Ser Pro Val Pro Thr 6515 6520 6525 Thr Leu Pro Glu Ser Arg Pro Ser Ser Pro Val His Leu Thr Ser 6530 6535 6540 Leu Pro Thr Ser Gly Leu Val Lys Thr Thr Asp Met Leu Ala Ser 6545 6550 6555 Val Ala Ser Leu Pro Pro Asn Leu Gly Ser Thr Ser His Lys Ile 6560 6565 6570 Pro Thr Thr Ser Glu Asp Ile Lys Asp Thr Glu Lys Met Tyr Pro 6575 6580 6585 Ser Thr Asn Ile Ala Val Thr Asn Val Gly Thr Thr Thr Ser Glu 6590 6595 6600 Lys Glu Ser Tyr Ser Ser Val Pro Ala Tyr Ser Glu Pro Pro Lys 6605 6610 6615 Val Thr Ser Pro Met Val Thr Ser Phe Asn Ile Arg Asp Thr Ile 6620 6625 6630 Val Ser Thr Ser Met Pro Gly Ser Ser Glu Ile Thr Arg Ile Glu 6635 6640 6645 Met Glu Ser Thr Phe Ser Leu Ala His Gly Leu Lys Gly Thr Ser 6650 6655 6660 Thr Ser Gln Asp Pro Ile Val Ser Thr Glu Lys Ser Ala Val Leu 6665 6670 6675 His Lys Leu Thr Thr Gly Ala Thr Glu Thr Ser Arg Thr Glu Val 6680 6685 6690 Ala Ser Ser Arg Arg Thr Ser Ile Pro Gly Pro Asp His Ser Thr 6695 6700 6705 Glu Ser Pro Asp Ile Ser Thr Glu Val Ile Pro Ser Leu Pro Ile 6710 6715 6720 Ser Leu Gly Ile Thr Glu Ser Ser Asn Met Thr Ile Ile Thr Arg 6725 6730 6735 Thr Gly Pro Pro Leu Gly Ser Thr Ser Gln Gly Thr Phe Thr Leu 6740 6745 6750 Asp Thr Pro Thr Thr Ser Ser Arg Ala Gly Thr His Ser Met Ala 6755 6760 6765 Thr Gln Glu Phe Pro His Ser Glu Met Thr Thr Val Met Asn Lys 6770 6775 6780 Asp Pro Glu Ile Leu Ser Trp Thr Ile Pro Pro Ser Ile Glu Lys 6785 6790 6795 Thr Ser Phe Ser Ser Ser Leu Met Pro Ser Pro Ala Met Thr Ser 6800 6805 6810 Pro Pro Val Ser Ser Thr Leu Pro Lys Thr Ile His Thr Thr Pro 6815 6820 6825 Ser Pro Met Thr Ser Leu Leu Thr Pro Ser Leu Val Met Thr Thr 6830 6835 6840 Asp Thr Leu Gly Thr Ser Pro Glu Pro Thr Thr Ser Ser Pro Pro 6845 6850 6855 Asn Leu Ser Ser Thr Ser His Glu Ile Leu Thr Thr Asp Glu Asp 6860 6865 6870 Thr Thr Ala Ile Glu Ala Met His Pro Ser Thr Ser Thr Ala Ala 6875 6880 6885 Thr Asn Val Glu Thr Thr Ser Ser Gly His Gly Ser Gln Ser Ser 6890 6895 6900 Val Leu Ala Asp Ser Glu Lys Thr Lys Ala Thr Ala Pro Met Asp 6905 6910 6915 Thr Thr Ser Thr Met Gly His Thr Thr Val Ser Thr Ser Met Ser 6920 6925 6930 Val Ser Ser Glu Thr Thr Lys Ile Lys Arg Glu Ser Thr Tyr Ser 6935 6940 6945 Leu Thr Pro Gly Leu Arg Glu Thr Ser Ile Ser Gln Asn Ala Ser 6950 6955 6960 Phe Ser Thr Asp Thr Ser Ile Val Leu Ser Glu Val Pro Thr Gly 6965 6970 6975 Thr Thr Ala Glu Val Ser Arg Thr Glu Val Thr Ser Ser Gly Arg 6980 6985 6990 Thr Ser Ile Pro Gly Pro Ser Gln Ser Thr Val Leu Pro Glu Ile 6995 7000 7005 Ser Thr Arg Thr Met Thr Arg Leu Phe Ala Ser Pro Thr Met Thr 7010 7015 7020 Glu Ser Ala Glu Met Thr Ile Pro Thr Gln Thr Gly Pro Ser Gly 7025 7030 7035 Ser Thr Ser Gln Asp Thr Leu Thr Leu Asp Thr Ser Thr Thr Lys 7040 7045 7050 Ser Gln Ala Lys Thr His Ser Thr Leu Thr Gln Arg Phe Pro His 7055 7060 7065 Ser Glu Met Thr Thr Leu Met Ser Arg Gly Pro Gly Asp Met Ser 7070 7075 7080 Trp Gln Ser Ser Pro Ser Leu Glu Asn Pro Ser Ser Leu Pro Ser 7085 7090 7095 Leu Leu Ser Leu Pro Ala Thr Thr Ser Pro Pro Pro Ile Ser Ser 7100 7105 7110 Thr Leu Pro Val Thr Ile Ser Ser Ser Pro Leu Pro Val Thr Ser 7115 7120 7125 Leu Leu Thr Ser Ser Pro Val Thr Thr Thr Asp Met Leu His Thr 7130 7135 7140 Ser Pro Glu Leu Val Thr Ser Ser Pro Pro Lys Leu Ser His Thr 7145 7150 7155 Ser Asp Glu Arg Leu Thr Thr Gly Lys Asp Thr Thr Asn Thr Glu 7160 7165 7170 Ala Val His Pro Ser Thr Asn Thr Ala Ala Ser Asn Val Glu Ile 7175 7180 7185 Pro Ser Ser Gly His Glu Ser Pro Ser Ser Ala Leu Ala Asp Ser 7190 7195 7200 Glu Thr Ser Lys Ala Thr Ser Pro Met Phe Ile Thr Ser Thr Gln 7205 7210 7215 Glu Asp Thr Thr Val Ala Ile Ser Thr Pro His Phe Leu Glu Thr 7220 7225 7230 Ser Arg Ile Gln Lys Glu Ser Ile Ser Ser Leu Ser Pro Lys Leu 7235 7240 7245 Arg Glu Thr Gly Ser Ser Val Glu Thr Ser Ser Ala Ile Glu Thr 7250 7255 7260 Ser Ala Val Leu Ser Glu Val Ser Ile Gly Ala Thr Thr Glu Ile 7265 7270 7275 Ser Arg Thr Glu Val Thr Ser Ser Ser Arg Thr Ser Ile Ser Gly 7280 7285 7290 Ser Ala Glu Ser Thr Met Leu Pro Glu Ile Ser Thr Thr Arg Lys 7295 7300 7305 Ile Ile Lys Phe Pro Thr Ser Pro Ile Leu Ala Glu Ser Ser Glu 7310 7315 7320 Met Thr Ile Lys Thr Gln Thr Ser Pro Pro Gly Ser Thr Ser Glu 7325 7330 7335 Ser Thr Phe Thr Leu Asp Thr Ser Thr Thr Pro Ser Leu Val Ile 7340 7345 7350 Thr His Ser Thr Met Thr Gln Arg Leu Pro His Ser Glu Ile Thr 7355 7360 7365 Thr Leu Val Ser Arg Gly Ala Gly Asp Val Pro Arg Pro Ser Ser 7370 7375 7380 Leu Pro Val Glu Glu Thr Ser Pro Pro Ser Ser Gln Leu Ser Leu 7385 7390 7395 Ser Ala Met Ile Ser Pro Ser Pro Val Ser Ser Thr Leu Pro Ala 7400 7405 7410 Ser Ser His Ser Ser Ser Ala Ser Val Thr Ser Leu Leu Thr Pro 7415 7420 7425 Gly Gln Val Lys Thr Thr Glu Val Leu Asp Ala Ser Ala Glu Pro 7430 7435 7440 Glu Thr Ser Ser Pro Pro Ser Leu Ser Ser Thr Ser Val Glu Ile 7445 7450 7455 Leu Ala Thr Ser Glu Val Thr Thr Asp Thr Glu Lys Ile His Pro 7460 7465 7470 Phe Ser Asn Thr Ala Val Thr Lys Val Gly Thr Ser Ser Ser Gly 7475 7480 7485 His Glu Ser Pro Ser Ser Val Leu Pro Asp Ser Glu Thr Thr Lys 7490 7495 7500 Ala Thr Ser Ala Met Gly Thr Ile Ser Ile Met Gly Asp Thr Ser 7505 7510 7515 Val Ser Thr Leu Thr Pro Ala Leu Ser Asn Thr Arg Lys Ile Gln 7520 7525 7530 Ser Glu Pro Ala Ser Ser Leu Thr Thr Arg Leu Arg Glu Thr Ser 7535 7540 7545 Thr Ser Glu Glu Thr Ser Leu Ala Thr Glu Ala Asn Thr Val Leu 7550 7555 7560 Ser Lys Val Ser Thr Gly Ala Thr Thr Glu Val Ser Arg Thr Glu 7565 7570 7575 Ala Ile Ser Phe Ser Arg Thr Ser Met Ser Gly Pro Glu Gln Ser 7580 7585 7590 Thr Met Ser Gln Asp Ile Ser Ile Gly Thr Ile Pro Arg Ile Ser 7595 7600 7605 Ala Ser Ser Val Leu Thr Glu Ser Ala Lys Met Thr Ile Thr Thr 7610 7615 7620 Gln Thr Gly Pro Ser Glu Ser Thr Leu Glu Ser Thr Leu Asn Leu 7625 7630 7635 Asn Thr Ala Thr Thr Pro Ser Trp Val Glu Thr His Ser Ile Val 7640 7645 7650 Ile Gln Gly Phe Pro His Pro Glu Met Thr Thr Ser Met Gly Arg 7655 7660 7665 Gly Pro Gly Gly Val Ser Trp Pro Ser Pro Pro Phe Val Lys Glu 7670 7675 7680 Thr Ser Pro Pro Ser Ser Pro Leu Ser Leu Pro Ala Val Thr Ser 7685 7690 7695 Pro His Pro Val Ser Thr Thr Phe Leu Ala His Ile Pro Pro Ser 7700 7705 7710 Pro Leu Pro Val Thr Ser Leu Leu Thr Ser Gly Pro Ala Thr Thr 7715 7720 7725 Thr Asp Ile Leu Gly Thr Ser Thr Glu Pro Gly Thr Ser Ser Ser 7730 7735 7740 Ser Ser Leu Ser Thr Thr Ser His Glu Arg Leu Thr Thr Tyr Lys 7745 7750 7755 Asp Thr Ala His Thr Glu Ala Val His Pro Ser Thr Asn Thr Gly 7760 7765 7770 Gly Thr Asn Val Ala Thr Thr Ser Ser Gly Tyr Lys Ser Gln Ser 7775 7780 7785 Ser Val Leu Ala Asp Ser Ser Pro Met Cys Thr Thr Ser Thr Met 7790 7795 7800 Gly Asp Thr Ser Val Leu Thr Ser Thr Pro Ala Phe Leu Glu Thr 7805 7810 7815 Arg Arg Ile Gln Thr Glu Leu Ala Ser Ser Leu Thr Pro Gly Leu 7820 7825 7830 Arg Glu Ser Ser Gly Ser Glu Gly Thr Ser Ser Gly Thr Lys Met 7835 7840 7845 Ser Thr Val Leu Ser Lys Val Pro Thr Gly Ala Thr Thr Glu Ile 7850 7855 7860 Ser Lys Glu Asp Val Thr Ser Ile Pro Gly Pro Ala Gln Ser Thr 7865 7870 7875 Ile Ser Pro Asp Ile Ser Thr Arg Thr Val Ser Trp Phe Ser Thr 7880 7885 7890 Ser Pro Val Met Thr Glu Ser Ala Glu Ile Thr Met Asn Thr His 7895 7900 7905 Thr Ser Pro Leu Gly Ala Thr Thr Gln Gly Thr Ser Thr Leu Asp 7910 7915 7920 Thr Ser Ser Thr Thr Ser Leu Thr Met Thr His Ser Thr Ile Ser 7925 7930 7935 Gln Gly Phe Ser His Ser Gln Met Ser Thr Leu Met Arg Arg Gly 7940 7945 7950 Pro Glu Asp Val Ser Trp Met Ser Pro Pro Leu Leu Glu Lys Thr 7955 7960 7965 Arg Pro Ser Phe Ser Leu Met Ser Ser Pro Ala Thr Thr Ser Pro 7970 7975 7980 Ser Pro Val Ser Ser Thr Leu Pro Glu Ser Ile Ser Ser Ser Pro 7985 7990 7995 Leu Pro Val Thr Ser Leu Leu Thr Ser Gly Leu Ala Lys Thr Thr 8000 8005 8010 Asp Met Leu His Lys Ser Ser Glu Pro Val Thr Asn Ser Pro Ala 8015 8020 8025 Asn Leu Ser Ser Thr Ser Val Glu Ile Leu Ala Thr Ser Glu Val 8030 8035 8040 Thr Thr Asp Thr Glu Lys Thr His Pro Ser Ser Asn Arg Thr Val 8045 8050 8055 Thr Asp Val Gly Thr Ser Ser Ser Gly His Glu Ser Thr Ser Phe 8060 8065 8070 Val Leu Ala Asp Ser Gln Thr Ser Lys Val Thr Ser Pro Met Val 8075 8080 8085 Ile Thr Ser Thr Met Glu Asp Thr Ser Val Ser Thr Ser Thr Pro 8090 8095 8100 Gly Phe Phe Glu Thr Ser Arg Ile Gln Thr Glu Pro Thr Ser Ser 8105 8110 8115 Leu Thr Leu Gly Leu Arg Lys Thr Ser Ser Ser Glu Gly Thr Ser 8120 8125 8130 Leu Ala Thr Glu Met Ser Thr Val Leu Ser Gly Val Pro Thr Gly 8135 8140 8145 Ala Thr Ala Glu Val Ser Arg Thr Glu Val Thr Ser Ser Ser Arg 8150 8155 8160 Thr Ser Ile Ser Gly Phe Ala Gln Leu Thr Val Ser Pro Glu Thr 8165 8170 8175 Ser Thr Glu Thr Ile Thr Arg Leu Pro Thr Ser Ser Ile Met Thr 8180 8185 8190 Glu Ser Ala Glu Met Met Ile Lys Thr Gln Thr Asp Pro Pro Gly 8195 8200 8205 Ser Thr Pro Glu Ser Thr His Thr Val Asp Ile Ser Thr Thr Pro 8210 8215 8220 Asn Trp Val Glu Thr His Ser Thr Val Thr Gln Arg Phe Ser His 8225 8230 8235 Ser Glu Met Thr Thr Leu Val Ser Arg Ser Pro Gly Asp Met Leu 8240 8245 8250 Trp Pro Ser Gln Ser Ser Val Glu Glu Thr Ser Ser Ala Ser Ser 8255 8260 8265 Leu Leu Ser Leu Pro Ala Thr Thr Ser Pro Ser Pro Val Ser Ser 8270 8275 8280 Thr Leu Val Glu Asp Phe Pro Ser Ala Ser Leu Pro Val Thr Ser 8285 8290 8295 Leu Leu Asn Pro Gly Leu Val Ile Thr Thr Asp Arg Met Gly Ile 8300 8305 8310 Ser Arg Glu Pro Gly Thr Ser Ser Thr Ser Asn Leu Ser Ser Thr 8315 8320 8325 Ser His Glu Arg Leu Thr Thr Leu Glu Asp Thr Val Asp Thr Glu 8330 8335 8340 Asp Met Gln Pro Ser Thr His Thr Ala Val Thr Asn Val Arg Thr 8345 8350 8355 Ser Ile Ser Gly His Glu Ser Gln Ser Ser Val Leu Ser Asp Ser 8360 8365 8370 Glu Thr Pro Lys Ala Thr Ser Pro Met Gly Thr Thr Tyr Thr Met 8375 8380 8385 Gly Glu Thr Ser Val Ser Ile Ser Thr Ser Asp Phe Phe Glu Thr 8390 8395 8400 Ser Arg Ile Gln Ile Glu Pro Thr Ser Ser Leu Thr Ser Gly Leu 8405 8410 8415 Arg Glu Thr Ser Ser Ser Glu Arg Ile Ser Ser Ala Thr Glu Gly 8420 8425 8430 Ser Thr Val Leu Ser Glu Val Pro Ser Gly Ala Thr Thr Glu Val 8435 8440 8445 Ser Arg Thr Glu Val Ile Ser Ser Arg Gly Thr Ser Met Ser Gly 8450

8455 8460 Pro Asp Gln Phe Thr Ile Ser Pro Asp Ile Ser Thr Glu Ala Ile 8465 8470 8475 Thr Arg Leu Ser Thr Ser Pro Ile Met Thr Glu Ser Ala Glu Ser 8480 8485 8490 Ala Ile Thr Ile Glu Thr Gly Ser Pro Gly Ala Thr Ser Glu Gly 8495 8500 8505 Thr Leu Thr Leu Asp Thr Ser Thr Thr Thr Phe Trp Ser Gly Thr 8510 8515 8520 His Ser Thr Ala Ser Pro Gly Phe Ser His Ser Glu Met Thr Thr 8525 8530 8535 Leu Met Ser Arg Thr Pro Gly Asp Val Pro Trp Pro Ser Leu Pro 8540 8545 8550 Ser Val Glu Glu Ala Ser Ser Val Ser Ser Ser Leu Ser Ser Pro 8555 8560 8565 Ala Met Thr Ser Thr Ser Phe Phe Ser Thr Leu Pro Glu Ser Ile 8570 8575 8580 Ser Ser Ser Pro His Pro Val Thr Ala Leu Leu Thr Leu Gly Pro 8585 8590 8595 Val Lys Thr Thr Asp Met Leu Arg Thr Ser Ser Glu Pro Glu Thr 8600 8605 8610 Ser Ser Pro Pro Asn Leu Ser Ser Thr Ser Ala Glu Ile Leu Ala 8615 8620 8625 Thr Ser Glu Val Thr Lys Asp Arg Glu Lys Ile His Pro Ser Ser 8630 8635 8640 Asn Thr Pro Val Val Asn Val Gly Thr Val Ile Tyr Lys His Leu 8645 8650 8655 Ser Pro Ser Ser Val Leu Ala Asp Leu Val Thr Thr Lys Pro Thr 8660 8665 8670 Ser Pro Met Ala Thr Thr Ser Thr Leu Gly Asn Thr Ser Val Ser 8675 8680 8685 Thr Ser Thr Pro Ala Phe Pro Glu Thr Met Met Thr Gln Pro Thr 8690 8695 8700 Ser Ser Leu Thr Ser Gly Leu Arg Glu Ile Ser Thr Ser Gln Glu 8705 8710 8715 Thr Ser Ser Ala Thr Glu Arg Ser Ala Ser Leu Ser Gly Met Pro 8720 8725 8730 Thr Gly Ala Thr Thr Lys Val Ser Arg Thr Glu Ala Leu Ser Leu 8735 8740 8745 Gly Arg Thr Ser Thr Pro Gly Pro Ala Gln Ser Thr Ile Ser Pro 8750 8755 8760 Glu Ile Ser Thr Glu Thr Ile Thr Arg Ile Ser Thr Pro Leu Thr 8765 8770 8775 Thr Thr Gly Ser Ala Glu Met Thr Ile Thr Pro Lys Thr Gly His 8780 8785 8790 Ser Gly Ala Ser Ser Gln Gly Thr Phe Thr Leu Asp Thr Ser Ser 8795 8800 8805 Arg Ala Ser Trp Pro Gly Thr His Ser Ala Ala Thr His Arg Ser 8810 8815 8820 Pro His Ser Gly Met Thr Thr Pro Met Ser Arg Gly Pro Glu Asp 8825 8830 8835 Val Ser Trp Pro Ser Arg Pro Ser Val Glu Lys Thr Ser Pro Pro 8840 8845 8850 Ser Ser Leu Val Ser Leu Ser Ala Val Thr Ser Pro Ser Pro Leu 8855 8860 8865 Tyr Ser Thr Pro Ser Glu Ser Ser His Ser Ser Pro Leu Arg Val 8870 8875 8880 Thr Ser Leu Phe Thr Pro Val Met Met Lys Thr Thr Asp Met Leu 8885 8890 8895 Asp Thr Ser Leu Glu Pro Val Thr Thr Ser Pro Pro Ser Met Asn 8900 8905 8910 Ile Thr Ser Asp Glu Ser Leu Ala Thr Ser Lys Ala Thr Met Glu 8915 8920 8925 Thr Glu Ala Ile Gln Leu Ser Glu Asn Thr Ala Val Thr Gln Met 8930 8935 8940 Gly Thr Ile Ser Ala Arg Gln Glu Phe Tyr Ser Ser Tyr Pro Gly 8945 8950 8955 Leu Pro Glu Pro Ser Lys Val Thr Ser Pro Val Val Thr Ser Ser 8960 8965 8970 Thr Ile Lys Asp Ile Val Ser Thr Thr Ile Pro Ala Ser Ser Glu 8975 8980 8985 Ile Thr Arg Ile Glu Met Glu Ser Thr Ser Thr Leu Thr Pro Thr 8990 8995 9000 Pro Arg Glu Thr Ser Thr Ser Gln Glu Ile His Ser Ala Thr Lys 9005 9010 9015 Pro Ser Thr Val Pro Tyr Lys Ala Leu Thr Ser Ala Thr Ile Glu 9020 9025 9030 Asp Ser Met Thr Gln Val Met Ser Ser Ser Arg Gly Pro Ser Pro 9035 9040 9045 Asp Gln Ser Thr Met Ser Gln Asp Ile Ser Thr Glu Val Ile Thr 9050 9055 9060 Arg Leu Ser Thr Ser Pro Ile Lys Thr Glu Ser Thr Glu Met Thr 9065 9070 9075 Ile Thr Thr Gln Thr Gly Ser Pro Gly Ala Thr Ser Arg Gly Thr 9080 9085 9090 Leu Thr Leu Asp Thr Ser Thr Thr Phe Met Ser Gly Thr His Ser 9095 9100 9105 Thr Ala Ser Gln Gly Phe Ser His Ser Gln Met Thr Ala Leu Met 9110 9115 9120 Ser Arg Thr Pro Gly Asp Val Pro Trp Leu Ser His Pro Ser Val 9125 9130 9135 Glu Glu Ala Ser Ser Ala Ser Phe Ser Leu Ser Ser Pro Val Met 9140 9145 9150 Thr Ser Ser Ser Pro Val Ser Ser Thr Leu Pro Asp Ser Ile His 9155 9160 9165 Ser Ser Ser Leu Pro Val Thr Ser Leu Leu Thr Ser Gly Leu Val 9170 9175 9180 Lys Thr Thr Glu Leu Leu Gly Thr Ser Ser Glu Pro Glu Thr Ser 9185 9190 9195 Ser Pro Pro Asn Leu Ser Ser Thr Ser Ala Glu Ile Leu Ala Ile 9200 9205 9210 Thr Glu Val Thr Thr Asp Thr Glu Lys Leu Glu Met Thr Asn Val 9215 9220 9225 Val Thr Ser Gly Tyr Thr His Glu Ser Pro Ser Ser Val Leu Ala 9230 9235 9240 Asp Ser Val Thr Thr Lys Ala Thr Ser Ser Met Gly Ile Thr Tyr 9245 9250 9255 Pro Thr Gly Asp Thr Asn Val Leu Thr Ser Thr Pro Ala Phe Ser 9260 9265 9270 Asp Thr Ser Arg Ile Gln Thr Lys Ser Lys Leu Ser Leu Thr Pro 9275 9280 9285 Gly Leu Met Glu Thr Ser Ile Ser Glu Glu Thr Ser Ser Ala Thr 9290 9295 9300 Glu Lys Ser Thr Val Leu Ser Ser Val Pro Thr Gly Ala Thr Thr 9305 9310 9315 Glu Val Ser Arg Thr Glu Ala Ile Ser Ser Ser Arg Thr Ser Ile 9320 9325 9330 Pro Gly Pro Ala Gln Ser Thr Met Ser Ser Asp Thr Ser Met Glu 9335 9340 9345 Thr Ile Thr Arg Ile Ser Thr Pro Leu Thr Arg Lys Glu Ser Thr 9350 9355 9360 Asp Met Ala Ile Thr Pro Lys Thr Gly Pro Ser Gly Ala Thr Ser 9365 9370 9375 Gln Gly Thr Phe Thr Leu Asp Ser Ser Ser Thr Ala Ser Trp Pro 9380 9385 9390 Gly Thr His Ser Ala Thr Thr Gln Arg Phe Pro Gln Ser Val Val 9395 9400 9405 Thr Thr Pro Met Ser Arg Gly Pro Glu Asp Val Ser Trp Pro Ser 9410 9415 9420 Pro Leu Ser Val Glu Lys Asn Ser Pro Pro Ser Ser Leu Val Ser 9425 9430 9435 Ser Ser Ser Val Thr Ser Pro Ser Pro Leu Tyr Ser Thr Pro Ser 9440 9445 9450 Gly Ser Ser His Ser Ser Pro Val Pro Val Thr Ser Leu Phe Thr 9455 9460 9465 Ser Ile Met Met Lys Ala Thr Asp Met Leu Asp Ala Ser Leu Glu 9470 9475 9480 Pro Glu Thr Thr Ser Ala Pro Asn Met Asn Ile Thr Ser Asp Glu 9485 9490 9495 Ser Leu Ala Ala Ser Lys Ala Thr Thr Glu Thr Glu Ala Ile His 9500 9505 9510 Val Phe Glu Asn Thr Ala Ala Ser His Val Glu Thr Thr Ser Ala 9515 9520 9525 Thr Glu Glu Leu Tyr Ser Ser Ser Pro Gly Phe Ser Glu Pro Thr 9530 9535 9540 Lys Val Ile Ser Pro Val Val Thr Ser Ser Ser Ile Arg Asp Asn 9545 9550 9555 Met Val Ser Thr Thr Met Pro Gly Ser Ser Gly Ile Thr Arg Ile 9560 9565 9570 Glu Ile Glu Ser Met Ser Ser Leu Thr Pro Gly Leu Arg Glu Thr 9575 9580 9585 Arg Thr Ser Gln Asp Ile Thr Ser Ser Thr Glu Thr Ser Thr Val 9590 9595 9600 Leu Tyr Lys Met Pro Ser Gly Ala Thr Pro Glu Val Ser Arg Thr 9605 9610 9615 Glu Val Met Pro Ser Ser Arg Thr Ser Ile Pro Gly Pro Ala Gln 9620 9625 9630 Ser Thr Met Ser Leu Asp Ile Ser Asp Glu Val Val Thr Arg Leu 9635 9640 9645 Ser Thr Ser Pro Ile Met Thr Glu Ser Ala Glu Ile Thr Ile Thr 9650 9655 9660 Thr Gln Thr Gly Tyr Ser Leu Ala Thr Ser Gln Val Thr Leu Pro 9665 9670 9675 Leu Gly Thr Ser Met Thr Phe Leu Ser Gly Thr His Ser Thr Met 9680 9685 9690 Ser Gln Gly Leu Ser His Ser Glu Met Thr Asn Leu Met Ser Arg 9695 9700 9705 Gly Pro Glu Ser Leu Ser Trp Thr Ser Pro Arg Phe Val Glu Thr 9710 9715 9720 Thr Arg Ser Ser Ser Ser Leu Thr Ser Leu Pro Leu Thr Thr Ser 9725 9730 9735 Leu Ser Pro Val Ser Ser Thr Leu Leu Asp Ser Ser Pro Ser Ser 9740 9745 9750 Pro Leu Pro Val Thr Ser Leu Ile Leu Pro Gly Leu Val Lys Thr 9755 9760 9765 Thr Glu Val Leu Asp Thr Ser Ser Glu Pro Lys Thr Ser Ser Ser 9770 9775 9780 Pro Asn Leu Ser Ser Thr Ser Val Glu Ile Pro Ala Thr Ser Glu 9785 9790 9795 Ile Met Thr Asp Thr Glu Lys Ile His Pro Ser Ser Asn Thr Ala 9800 9805 9810 Val Ala Lys Val Arg Thr Ser Ser Ser Val His Glu Ser His Ser 9815 9820 9825 Ser Val Leu Ala Asp Ser Glu Thr Thr Ile Thr Ile Pro Ser Met 9830 9835 9840 Gly Ile Thr Ser Ala Val Asp Asp Thr Thr Val Phe Thr Ser Asn 9845 9850 9855 Pro Ala Phe Ser Glu Thr Arg Arg Ile Pro Thr Glu Pro Thr Phe 9860 9865 9870 Ser Leu Thr Pro Gly Phe Arg Glu Thr Ser Thr Ser Glu Glu Thr 9875 9880 9885 Thr Ser Ile Thr Glu Thr Ser Ala Val Leu Tyr Gly Val Pro Thr 9890 9895 9900 Ser Ala Thr Thr Glu Val Ser Met Thr Glu Ile Met Ser Ser Asn 9905 9910 9915 Arg Ile His Ile Pro Asp Ser Asp Gln Ser Thr Met Ser Pro Asp 9920 9925 9930 Ile Ile Thr Glu Val Ile Thr Arg Leu Ser Ser Ser Ser Met Met 9935 9940 9945 Ser Glu Ser Thr Gln Met Thr Ile Thr Thr Gln Lys Ser Ser Pro 9950 9955 9960 Gly Ala Thr Ala Gln Ser Thr Leu Thr Leu Ala Thr Thr Thr Ala 9965 9970 9975 Pro Leu Ala Arg Thr His Ser Thr Val Pro Pro Arg Phe Leu His 9980 9985 9990 Ser Glu Met Thr Thr Leu Met Ser Arg Ser Pro Glu Asn Pro Ser 9995 10000 10005 Trp Lys Ser Ser Leu Phe Val Glu Lys Thr Ser Ser Ser Ser Ser 10010 10015 10020 Leu Leu Ser Leu Pro Val Thr Thr Ser Pro Ser Val Ser Ser Thr 10025 10030 10035 Leu Pro Gln Ser Ile Pro Ser Ser Ser Phe Ser Val Thr Ser Leu 10040 10045 10050 Leu Thr Pro Gly Met Val Lys Thr Thr Asp Thr Ser Thr Glu Pro 10055 10060 10065 Gly Thr Ser Leu Ser Pro Asn Leu Ser Gly Thr Ser Val Glu Ile 10070 10075 10080 Leu Ala Ala Ser Glu Val Thr Thr Asp Thr Glu Lys Ile His Pro 10085 10090 10095 Ser Ser Ser Met Ala Val Thr Asn Val Gly Thr Thr Ser Ser Gly 10100 10105 10110 His Glu Leu Tyr Ser Ser Val Ser Ile His Ser Glu Pro Ser Lys 10115 10120 10125 Ala Thr Tyr Pro Val Gly Thr Pro Ser Ser Met Ala Glu Thr Ser 10130 10135 10140 Ile Ser Thr Ser Met Pro Ala Asn Phe Glu Thr Thr Gly Phe Glu 10145 10150 10155 Ala Glu Pro Phe Ser His Leu Thr Ser Gly Phe Arg Lys Thr Asn 10160 10165 10170 Met Ser Leu Asp Thr Ser Ser Val Thr Pro Thr Asn Thr Pro Ser 10175 10180 10185 Ser Pro Gly Ser Thr His Leu Leu Gln Ser Ser Lys Thr Asp Phe 10190 10195 10200 Thr Ser Ser Ala Lys Thr Ser Ser Pro Asp Trp Pro Pro Ala Ser 10205 10210 10215 Gln Tyr Thr Glu Ile Pro Val Asp Ile Ile Thr Pro Phe Asn Ala 10220 10225 10230 Ser Pro Ser Ile Thr Glu Ser Thr Gly Ile Thr Ser Phe Pro Glu 10235 10240 10245 Ser Arg Phe Thr Met Ser Val Thr Glu Ser Thr His His Leu Ser 10250 10255 10260 Thr Asp Leu Leu Pro Ser Ala Glu Thr Ile Ser Thr Gly Thr Val 10265 10270 10275 Met Pro Ser Leu Ser Glu Ala Met Thr Ser Phe Ala Thr Thr Gly 10280 10285 10290 Val Pro Arg Ala Ile Ser Gly Ser Gly Ser Pro Phe Ser Arg Thr 10295 10300 10305 Glu Ser Gly Pro Gly Asp Ala Thr Leu Ser Thr Ile Ala Glu Ser 10310 10315 10320 Leu Pro Ser Ser Thr Pro Val Pro Phe Ser Ser Ser Thr Phe Thr 10325 10330 10335 Thr Thr Asp Ser Ser Thr Ile Pro Ala Leu His Glu Ile Thr Ser 10340 10345 10350 Ser Ser Ala Thr Pro Tyr Arg Val Asp Thr Ser Leu Gly Thr Glu 10355 10360 10365 Ser Ser Thr Thr Glu Gly Arg Leu Val Met Val Ser Thr Leu Asp 10370 10375 10380 Thr Ser Ser Gln Pro Gly Arg Thr Ser Ser Ser Pro Ile Leu Asp 10385 10390 10395 Thr Arg Met Thr Glu Ser Val Glu Leu Gly Thr Val Thr Ser Ala 10400 10405 10410 Tyr Gln Val Pro Ser Leu Ser Thr Arg Leu Thr Arg Thr Asp Gly 10415 10420 10425 Ile Met Glu His Ile Thr Lys Ile Pro Asn Glu Ala Ala His Arg 10430 10435 10440 Gly Thr Ile Arg Pro Val Lys Gly Pro Gln Thr Ser Thr Ser Pro 10445 10450 10455 Ala Ser Pro Lys Gly Leu His Thr Gly Gly Thr Lys Arg Met Glu 10460 10465 10470 Thr Thr Thr Thr Ala Leu Lys Thr Thr Thr Thr Ala Leu Lys Thr 10475 10480 10485 Thr Ser Arg Ala Thr Leu Thr Thr Ser Val Tyr Thr Pro Thr Leu 10490 10495 10500 Gly Thr Leu Thr Pro Leu Asn Ala Ser Met Gln Met Ala Ser Thr 10505 10510 10515 Ile Pro Thr Glu Met Met Ile Thr Thr Pro Tyr Val Phe Pro Asp 10520 10525 10530 Val Pro Glu Thr Thr Ser Ser Leu Ala Thr Ser Leu Gly Ala Glu 10535 10540 10545 Thr Ser Thr Ala Leu Pro Arg Thr Thr Pro Ser Val Phe Asn Arg 10550 10555 10560 Glu Ser Glu Thr Thr Ala Ser Leu Val Ser Arg Ser Gly Ala Glu 10565 10570 10575 Arg Ser Pro Val Ile Gln Thr Leu Asp Val Ser Ser Ser Glu Pro 10580 10585 10590 Asp Thr Thr Ala Ser Trp Val Ile His Pro Ala Glu Thr Ile Pro 10595 10600 10605 Thr Val Ser Lys Thr Thr Pro Asn Phe Phe His Ser Glu Leu Asp 10610 10615 10620

Thr Val Ser Ser Thr Ala Thr Ser His Gly Ala Asp Val Ser Ser 10625 10630 10635 Ala Ile Pro Thr Asn Ile Ser Pro Ser Glu Leu Asp Ala Leu Thr 10640 10645 10650 Pro Leu Val Thr Ile Ser Gly Thr Asp Thr Ser Thr Thr Phe Pro 10655 10660 10665 Thr Leu Thr Lys Ser Pro His Glu Thr Glu Thr Arg Thr Thr Trp 10670 10675 10680 Leu Thr His Pro Ala Glu Thr Ser Ser Thr Ile Pro Arg Thr Ile 10685 10690 10695 Pro Asn Phe Ser His His Glu Ser Asp Ala Thr Pro Ser Ile Ala 10700 10705 10710 Thr Ser Pro Gly Ala Glu Thr Ser Ser Ala Ile Pro Ile Met Thr 10715 10720 10725 Val Ser Pro Gly Ala Glu Asp Leu Val Thr Ser Gln Val Thr Ser 10730 10735 10740 Ser Gly Thr Asp Arg Asn Met Thr Ile Pro Thr Leu Thr Leu Ser 10745 10750 10755 Pro Gly Glu Pro Lys Thr Ile Ala Ser Leu Val Thr His Pro Glu 10760 10765 10770 Ala Gln Thr Ser Ser Ala Ile Pro Thr Ser Thr Ile Ser Pro Ala 10775 10780 10785 Val Ser Arg Leu Val Thr Ser Met Val Thr Ser Leu Ala Ala Lys 10790 10795 10800 Thr Ser Thr Thr Asn Arg Ala Leu Thr Asn Ser Pro Gly Glu Pro 10805 10810 10815 Ala Thr Thr Val Ser Leu Val Thr His Pro Ala Gln Thr Ser Pro 10820 10825 10830 Thr Val Pro Trp Thr Thr Ser Ile Phe Phe His Ser Lys Ser Asp 10835 10840 10845 Thr Thr Pro Ser Met Thr Thr Ser His Gly Ala Glu Ser Ser Ser 10850 10855 10860 Ala Val Pro Thr Pro Thr Val Ser Thr Glu Val Pro Gly Val Val 10865 10870 10875 Thr Pro Leu Val Thr Ser Ser Arg Ala Val Ile Ser Thr Thr Ile 10880 10885 10890 Pro Ile Leu Thr Leu Ser Pro Gly Glu Pro Glu Thr Thr Pro Ser 10895 10900 10905 Met Ala Thr Ser His Gly Glu Glu Ala Ser Ser Ala Ile Pro Thr 10910 10915 10920 Pro Thr Val Ser Pro Gly Val Pro Gly Val Val Thr Ser Leu Val 10925 10930 10935 Thr Ser Ser Arg Ala Val Thr Ser Thr Thr Ile Pro Ile Leu Thr 10940 10945 10950 Phe Ser Leu Gly Glu Pro Glu Thr Thr Pro Ser Met Ala Thr Ser 10955 10960 10965 His Gly Thr Glu Ala Gly Ser Ala Val Pro Thr Val Leu Pro Glu 10970 10975 10980 Val Pro Gly Met Val Thr Ser Leu Val Ala Ser Ser Arg Ala Val 10985 10990 10995 Thr Ser Thr Thr Leu Pro Thr Leu Thr Leu Ser Pro Gly Glu Pro 11000 11005 11010 Glu Thr Thr Pro Ser Met Ala Thr Ser His Gly Ala Glu Ala Ser 11015 11020 11025 Ser Thr Val Pro Thr Val Ser Pro Glu Val Pro Gly Val Val Thr 11030 11035 11040 Ser Leu Val Thr Ser Ser Ser Gly Val Asn Ser Thr Ser Ile Pro 11045 11050 11055 Thr Leu Ile Leu Ser Pro Gly Glu Leu Glu Thr Thr Pro Ser Met 11060 11065 11070 Ala Thr Ser His Gly Ala Glu Ala Ser Ser Ala Val Pro Thr Pro 11075 11080 11085 Thr Val Ser Pro Gly Val Ser Gly Val Val Thr Pro Leu Val Thr 11090 11095 11100 Ser Ser Arg Ala Val Thr Ser Thr Thr Ile Pro Ile Leu Thr Leu 11105 11110 11115 Ser Ser Ser Glu Pro Glu Thr Thr Pro Ser Met Ala Thr Ser His 11120 11125 11130 Gly Val Glu Ala Ser Ser Ala Val Leu Thr Val Ser Pro Glu Val 11135 11140 11145 Pro Gly Met Val Thr Ser Leu Val Thr Ser Ser Arg Ala Val Thr 11150 11155 11160 Ser Thr Thr Ile Pro Thr Leu Thr Ile Ser Ser Asp Glu Pro Glu 11165 11170 11175 Thr Thr Thr Ser Leu Val Thr His Ser Glu Ala Lys Met Ile Ser 11180 11185 11190 Ala Ile Pro Thr Leu Ala Val Ser Pro Thr Val Gln Gly Leu Val 11195 11200 11205 Thr Ser Leu Val Thr Ser Ser Gly Ser Glu Thr Ser Ala Phe Ser 11210 11215 11220 Asn Leu Thr Val Ala Ser Ser Gln Pro Glu Thr Ile Asp Ser Trp 11225 11230 11235 Val Ala His Pro Gly Thr Glu Ala Ser Ser Val Val Pro Thr Leu 11240 11245 11250 Thr Val Ser Thr Gly Glu Pro Phe Thr Asn Ile Ser Leu Val Thr 11255 11260 11265 His Pro Ala Glu Ser Ser Ser Thr Leu Pro Arg Thr Thr Ser Arg 11270 11275 11280 Phe Ser His Ser Glu Leu Asp Thr Met Pro Ser Thr Val Thr Ser 11285 11290 11295 Pro Glu Ala Glu Ser Ser Ser Ala Ile Ser Thr Thr Ile Ser Pro 11300 11305 11310 Gly Ile Pro Gly Val Leu Thr Ser Leu Val Thr Ser Ser Gly Arg 11315 11320 11325 Asp Ile Ser Ala Thr Phe Pro Thr Val Pro Glu Ser Pro His Glu 11330 11335 11340 Ser Glu Ala Thr Ala Ser Trp Val Thr His Pro Ala Val Thr Ser 11345 11350 11355 Thr Thr Val Pro Arg Thr Thr Pro Asn Tyr Ser His Ser Glu Pro 11360 11365 11370 Asp Thr Thr Pro Ser Ile Ala Thr Ser Pro Gly Ala Glu Ala Thr 11375 11380 11385 Ser Asp Phe Pro Thr Ile Thr Val Ser Pro Asp Val Pro Asp Met 11390 11395 11400 Val Thr Ser Gln Val Thr Ser Ser Gly Thr Asp Thr Ser Ile Thr 11405 11410 11415 Ile Pro Thr Leu Thr Leu Ser Ser Gly Glu Pro Glu Thr Thr Thr 11420 11425 11430 Ser Phe Ile Thr Tyr Ser Glu Thr His Thr Ser Ser Ala Ile Pro 11435 11440 11445 Thr Leu Pro Val Ser Pro Gly Ala Ser Lys Met Leu Thr Ser Leu 11450 11455 11460 Val Ile Ser Ser Gly Thr Asp Ser Thr Thr Thr Phe Pro Thr Leu 11465 11470 11475 Thr Glu Thr Pro Tyr Glu Pro Glu Thr Thr Ala Ile Gln Leu Ile 11480 11485 11490 His Pro Ala Glu Thr Asn Thr Met Val Pro Arg Thr Thr Pro Lys 11495 11500 11505 Phe Ser His Ser Lys Ser Asp Thr Thr Leu Pro Val Ala Ile Thr 11510 11515 11520 Ser Pro Gly Pro Glu Ala Ser Ser Ala Val Ser Thr Thr Thr Ile 11525 11530 11535 Ser Pro Asp Met Ser Asp Leu Val Thr Ser Leu Val Pro Ser Ser 11540 11545 11550 Gly Thr Asp Thr Ser Thr Thr Phe Pro Thr Leu Ser Glu Thr Pro 11555 11560 11565 Tyr Glu Pro Glu Thr Thr Ala Thr Trp Leu Thr His Pro Ala Glu 11570 11575 11580 Thr Ser Thr Thr Val Ser Gly Thr Ile Pro Asn Phe Ser His Arg 11585 11590 11595 Gly Ser Asp Thr Ala Pro Ser Met Val Thr Ser Pro Gly Val Asp 11600 11605 11610 Thr Arg Ser Gly Val Pro Thr Thr Thr Ile Pro Pro Ser Ile Pro 11615 11620 11625 Gly Val Val Thr Ser Gln Val Thr Ser Ser Ala Thr Asp Thr Ser 11630 11635 11640 Thr Ala Ile Pro Thr Leu Thr Pro Ser Pro Gly Glu Pro Glu Thr 11645 11650 11655 Thr Ala Ser Ser Ala Thr His Pro Gly Thr Gln Thr Gly Phe Thr 11660 11665 11670 Val Pro Ile Arg Thr Val Pro Ser Ser Glu Pro Asp Thr Met Ala 11675 11680 11685 Ser Trp Val Thr His Pro Pro Gln Thr Ser Thr Pro Val Ser Arg 11690 11695 11700 Thr Thr Ser Ser Phe Ser His Ser Ser Pro Asp Ala Thr Pro Val 11705 11710 11715 Met Ala Thr Ser Pro Arg Thr Glu Ala Ser Ser Ala Val Leu Thr 11720 11725 11730 Thr Ile Ser Pro Gly Ala Pro Glu Met Val Thr Ser Gln Ile Thr 11735 11740 11745 Ser Ser Gly Ala Ala Thr Ser Thr Thr Val Pro Thr Leu Thr His 11750 11755 11760 Ser Pro Gly Met Pro Glu Thr Thr Ala Leu Leu Ser Thr His Pro 11765 11770 11775 Arg Thr Glu Thr Ser Lys Thr Phe Pro Ala Ser Thr Val Phe Pro 11780 11785 11790 Gln Val Ser Glu Thr Thr Ala Ser Leu Thr Ile Arg Pro Gly Ala 11795 11800 11805 Glu Thr Ser Thr Ala Leu Pro Thr Gln Thr Thr Ser Ser Leu Phe 11810 11815 11820 Thr Leu Leu Val Thr Gly Thr Ser Arg Val Asp Leu Ser Pro Thr 11825 11830 11835 Ala Ser Pro Gly Val Ser Ala Lys Thr Ala Pro Leu Ser Thr His 11840 11845 11850 Pro Gly Thr Glu Thr Ser Thr Met Ile Pro Thr Ser Thr Leu Ser 11855 11860 11865 Leu Gly Leu Leu Glu Thr Thr Gly Leu Leu Ala Thr Ser Ser Ser 11870 11875 11880 Ala Glu Thr Ser Thr Ser Thr Leu Thr Leu Thr Val Ser Pro Ala 11885 11890 11895 Val Ser Gly Leu Ser Ser Ala Ser Ile Thr Thr Asp Lys Pro Gln 11900 11905 11910 Thr Val Thr Ser Trp Asn Thr Glu Thr Ser Pro Ser Val Thr Ser 11915 11920 11925 Val Gly Pro Pro Glu Phe Ser Arg Thr Val Thr Gly Thr Thr Met 11930 11935 11940 Thr Leu Ile Pro Ser Glu Met Pro Thr Pro Pro Lys Thr Ser His 11945 11950 11955 Gly Glu Gly Val Ser Pro Thr Thr Ile Leu Arg Thr Thr Met Val 11960 11965 11970 Glu Ala Thr Asn Leu Ala Thr Thr Gly Ser Ser Pro Thr Val Ala 11975 11980 11985 Lys Thr Thr Thr Thr Phe Asn Thr Leu Ala Gly Ser Leu Phe Thr 11990 11995 12000 Pro Leu Thr Thr Pro Gly Met Ser Thr Leu Ala Ser Glu Ser Val 12005 12010 12015 Thr Ser Arg Thr Ser Tyr Asn His Arg Ser Trp Ile Ser Thr Thr 12020 12025 12030 Ser Ser Tyr Asn Arg Arg Tyr Trp Thr Pro Ala Thr Ser Thr Pro 12035 12040 12045 Val Thr Ser Thr Phe Ser Pro Gly Ile Ser Thr Ser Ser Ile Pro 12050 12055 12060 Ser Ser Thr Ala Ala Thr Val Pro Phe Met Val Pro Phe Thr Leu 12065 12070 12075 Asn Phe Thr Ile Thr Asn Leu Gln Tyr Glu Glu Asp Met Arg His 12080 12085 12090 Pro Gly Ser Arg Lys Phe Asn Ala Thr Glu Arg Glu Leu Gln Gly 12095 12100 12105 Leu Leu Lys Pro Leu Phe Arg Asn Ser Ser Leu Glu Tyr Leu Tyr 12110 12115 12120 Ser Gly Cys Arg Leu Ala Ser Leu Arg Pro Glu Lys Asp Ser Ser 12125 12130 12135 Ala Thr Ala Val Asp Ala Ile Cys Thr His Arg Pro Asp Pro Glu 12140 12145 12150 Asp Leu Gly Leu Asp Arg Glu Arg Leu Tyr Trp Glu Leu Ser Asn 12155 12160 12165 Leu Thr Asn Gly Ile Gln Glu Leu Gly Pro Tyr Thr Leu Asp Arg 12170 12175 12180 Asn Ser Leu Tyr Val Asn Gly Phe Thr His Arg Ser Ser Met Pro 12185 12190 12195 Thr Thr Ser Thr Pro Gly Thr Ser Thr Val Asp Val Gly Thr Ser 12200 12205 12210 Gly Thr Pro Ser Ser Ser Pro Ser Pro Thr Thr Ala Gly Pro Leu 12215 12220 12225 Leu Met Pro Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Gln Tyr 12230 12235 12240 Glu Glu Asp Met Arg Arg Thr Gly Ser Arg Lys Phe Asn Thr Met 12245 12250 12255 Glu Ser Val Leu Gln Gly Leu Leu Lys Pro Leu Phe Lys Asn Thr 12260 12265 12270 Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg 12275 12280 12285 Pro Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Ala Ile Cys Thr 12290 12295 12300 His Arg Leu Asp Pro Lys Ser Pro Gly Leu Asn Arg Glu Gln Leu 12305 12310 12315 Tyr Trp Glu Leu Ser Lys Leu Thr Asn Asp Ile Glu Glu Leu Gly 12320 12325 12330 Pro Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val Asn Gly Phe Thr 12335 12340 12345 His Gln Ser Ser Val Ser Thr Thr Ser Thr Pro Gly Thr Ser Thr 12350 12355 12360 Val Asp Leu Arg Thr Ser Gly Thr Pro Ser Ser Leu Ser Ser Pro 12365 12370 12375 Thr Ile Met Ala Ala Gly Pro Leu Leu Val Pro Phe Thr Leu Asn 12380 12385 12390 Phe Thr Ile Thr Asn Leu Gln Tyr Gly Glu Asp Met Gly His Pro 12395 12400 12405 Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg Val Leu Gln Gly Leu 12410 12415 12420 Leu Gly Pro Ile Phe Lys Asn Thr Ser Val Gly Pro Leu Tyr Ser 12425 12430 12435 Gly Cys Arg Leu Thr Ser Leu Arg Ser Glu Lys Asp Gly Ala Ala 12440 12445 12450 Thr Gly Val Asp Ala Ile Cys Ile His His Leu Asp Pro Lys Ser 12455 12460 12465 Pro Gly Leu Asn Arg Glu Arg Leu Tyr Trp Glu Leu Ser Gln Leu 12470 12475 12480 Thr Asn Gly Ile Lys Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asn 12485 12490 12495 Ser Leu Tyr Val Asn Gly Phe Thr His Arg Thr Ser Val Pro Thr 12500 12505 12510 Ser Ser Thr Pro Gly Thr Ser Thr Val Asp Leu Gly Thr Ser Gly 12515 12520 12525 Thr Pro Phe Ser Leu Pro Ser Pro Ala Thr Ala Gly Pro Leu Leu 12530 12535 12540 Val Leu Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Lys Tyr Glu 12545 12550 12555 Glu Asp Met His Arg Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu 12560 12565 12570 Arg Val Leu Gln Thr Leu Leu Gly Pro Met Phe Lys Asn Thr Ser 12575 12580 12585 Val Gly Leu Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Ser 12590 12595 12600 Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Ala Ile Cys Thr His 12605 12610 12615 Arg Leu Asp Pro Lys Ser Pro Gly Val Asp Arg Glu Gln Leu Tyr 12620 12625 12630 Trp Glu Leu Ser Gln Leu Thr Asn Gly Ile Lys Glu Leu Gly Pro 12635 12640 12645 Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val Asn Gly Phe Thr His 12650 12655 12660 Trp Ile Pro Val Pro Thr Ser Ser Thr Pro Gly Thr Ser Thr Val 12665 12670 12675 Asp Leu Gly Ser Gly Thr Pro Ser Ser Leu Pro Ser Pro Thr Thr 12680 12685 12690 Ala Gly Pro Leu Leu Val Pro Phe Thr Leu Asn Phe Thr Ile Thr 12695 12700 12705 Asn Leu Lys Tyr Glu Glu Asp Met His Cys Pro

Gly Ser Arg Lys 12710 12715 12720 Phe Asn Thr Thr Glu Arg Val Leu Gln Ser Leu Leu Gly Pro Met 12725 12730 12735 Phe Lys Asn Thr Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu 12740 12745 12750 Thr Leu Leu Arg Ser Glu Lys Asp Gly Ala Ala Thr Gly Val Asp 12755 12760 12765 Ala Ile Cys Thr His Arg Leu Asp Pro Lys Ser Pro Gly Val Asp 12770 12775 12780 Arg Glu Gln Leu Tyr Trp Glu Leu Ser Gln Leu Thr Asn Gly Ile 12785 12790 12795 Lys Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val 12800 12805 12810 Asn Gly Phe Thr His Gln Thr Ser Ala Pro Asn Thr Ser Thr Pro 12815 12820 12825 Gly Thr Ser Thr Val Asp Leu Gly Thr Ser Gly Thr Pro Ser Ser 12830 12835 12840 Leu Pro Ser Pro Thr Ser Ala Gly Pro Leu Leu Val Pro Phe Thr 12845 12850 12855 Leu Asn Phe Thr Ile Thr Asn Leu Gln Tyr Glu Glu Asp Met His 12860 12865 12870 His Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg Val Leu Gln 12875 12880 12885 Gly Leu Leu Gly Pro Met Phe Lys Asn Thr Ser Val Gly Leu Leu 12890 12895 12900 Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Pro Glu Lys Asn Gly 12905 12910 12915 Ala Ala Thr Gly Met Asp Ala Ile Cys Ser His Arg Leu Asp Pro 12920 12925 12930 Lys Ser Pro Gly Leu Asn Arg Glu Gln Leu Tyr Trp Glu Leu Ser 12935 12940 12945 Gln Leu Thr His Gly Ile Lys Glu Leu Gly Pro Tyr Thr Leu Asp 12950 12955 12960 Arg Asn Ser Leu Tyr Val Asn Gly Phe Thr His Arg Ser Ser Val 12965 12970 12975 Ala Pro Thr Ser Thr Pro Gly Thr Ser Thr Val Asp Leu Gly Thr 12980 12985 12990 Ser Gly Thr Pro Ser Ser Leu Pro Ser Pro Thr Thr Ala Val Pro 12995 13000 13005 Leu Leu Val Pro Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Gln 13010 13015 13020 Tyr Gly Glu Asp Met Arg His Pro Gly Ser Arg Lys Phe Asn Thr 13025 13030 13035 Thr Glu Arg Val Leu Gln Gly Leu Leu Gly Pro Leu Phe Lys Asn 13040 13045 13050 Ser Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu Ile Ser Leu 13055 13060 13065 Arg Ser Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Ala Ile Cys 13070 13075 13080 Thr His His Leu Asn Pro Gln Ser Pro Gly Leu Asp Arg Glu Gln 13085 13090 13095 Leu Tyr Trp Gln Leu Ser Gln Met Thr Asn Gly Ile Lys Glu Leu 13100 13105 13110 Gly Pro Tyr Thr Leu Asp Arg Asn Ser Leu Tyr Val Asn Gly Phe 13115 13120 13125 Thr His Arg Ser Ser Gly Leu Thr Thr Ser Thr Pro Trp Thr Ser 13130 13135 13140 Thr Val Asp Leu Gly Thr Ser Gly Thr Pro Ser Pro Val Pro Ser 13145 13150 13155 Pro Thr Thr Thr Gly Pro Leu Leu Val Pro Phe Thr Leu Asn Phe 13160 13165 13170 Thr Ile Thr Asn Leu Gln Tyr Glu Glu Asn Met Gly His Pro Gly 13175 13180 13185 Ser Arg Lys Phe Asn Ile Thr Glu Ser Val Leu Gln Gly Leu Leu 13190 13195 13200 Lys Pro Leu Phe Lys Ser Thr Ser Val Gly Pro Leu Tyr Ser Gly 13205 13210 13215 Cys Arg Leu Thr Leu Leu Arg Pro Glu Lys Asp Gly Val Ala Thr 13220 13225 13230 Arg Val Asp Ala Ile Cys Thr His Arg Pro Asp Pro Lys Ile Pro 13235 13240 13245 Gly Leu Asp Arg Gln Gln Leu Tyr Trp Glu Leu Ser Gln Leu Thr 13250 13255 13260 His Ser Ile Thr Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asp Ser 13265 13270 13275 Leu Tyr Val Asn Gly Phe Thr Gln Arg Ser Ser Val Pro Thr Thr 13280 13285 13290 Ser Thr Pro Gly Thr Phe Thr Val Gln Pro Glu Thr Ser Glu Thr 13295 13300 13305 Pro Ser Ser Leu Pro Gly Pro Thr Ala Thr Gly Pro Val Leu Leu 13310 13315 13320 Pro Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Gln Tyr Glu Glu 13325 13330 13335 Asp Met Arg Arg Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg 13340 13345 13350 Val Leu Gln Gly Leu Leu Met Pro Leu Phe Lys Asn Thr Ser Val 13355 13360 13365 Ser Ser Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Pro Glu 13370 13375 13380 Lys Asp Gly Ala Ala Thr Arg Val Asp Ala Val Cys Thr His Arg 13385 13390 13395 Pro Asp Pro Lys Ser Pro Gly Leu Asp Arg Glu Arg Leu Tyr Trp 13400 13405 13410 Lys Leu Ser Gln Leu Thr His Gly Ile Thr Glu Leu Gly Pro Tyr 13415 13420 13425 Thr Leu Asp Arg His Ser Leu Tyr Val Asn Gly Phe Thr His Gln 13430 13435 13440 Ser Ser Met Thr Thr Thr Arg Thr Pro Asp Thr Ser Thr Met His 13445 13450 13455 Leu Ala Thr Ser Arg Thr Pro Ala Ser Leu Ser Gly Pro Met Thr 13460 13465 13470 Ala Ser Pro Leu Leu Val Leu Phe Thr Ile Asn Phe Thr Ile Thr 13475 13480 13485 Asn Leu Arg Tyr Glu Glu Asn Met His His Pro Gly Ser Arg Lys 13490 13495 13500 Phe Asn Thr Thr Glu Arg Val Leu Gln Gly Leu Leu Arg Pro Val 13505 13510 13515 Phe Lys Asn Thr Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu 13520 13525 13530 Thr Leu Leu Arg Pro Lys Lys Asp Gly Ala Ala Thr Lys Val Asp 13535 13540 13545 Ala Ile Cys Thr Tyr Arg Pro Asp Pro Lys Ser Pro Gly Leu Asp 13550 13555 13560 Arg Glu Gln Leu Tyr Trp Glu Leu Ser Gln Leu Thr His Ser Ile 13565 13570 13575 Thr Glu Leu Gly Pro Tyr Thr Leu Asp Arg Asp Ser Leu Tyr Val 13580 13585 13590 Asn Gly Phe Thr Gln Arg Ser Ser Val Pro Thr Thr Ser Ile Pro 13595 13600 13605 Gly Thr Pro Thr Val Asp Leu Gly Thr Ser Gly Thr Pro Val Ser 13610 13615 13620 Lys Pro Gly Pro Ser Ala Ala Ser Pro Leu Leu Val Leu Phe Thr 13625 13630 13635 Leu Asn Phe Thr Ile Thr Asn Leu Arg Tyr Glu Glu Asn Met Gln 13640 13645 13650 His Pro Gly Ser Arg Lys Phe Asn Thr Thr Glu Arg Val Leu Gln 13655 13660 13665 Gly Leu Leu Arg Ser Leu Phe Lys Ser Thr Ser Val Gly Pro Leu 13670 13675 13680 Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg Pro Glu Lys Asp Gly 13685 13690 13695 Thr Ala Thr Gly Val Asp Ala Ile Cys Thr His His Pro Asp Pro 13700 13705 13710 Lys Ser Pro Arg Leu Asp Arg Glu Gln Leu Tyr Trp Glu Leu Ser 13715 13720 13725 Gln Leu Thr His Asn Ile Thr Glu Leu Gly Pro Tyr Ala Leu Asp 13730 13735 13740 Asn Asp Ser Leu Phe Val Asn Gly Phe Thr His Arg Ser Ser Val 13745 13750 13755 Ser Thr Thr Ser Thr Pro Gly Thr Pro Thr Val Tyr Leu Gly Ala 13760 13765 13770 Ser Lys Thr Pro Ala Ser Ile Phe Gly Pro Ser Ala Ala Ser His 13775 13780 13785 Leu Leu Ile Leu Phe Thr Leu Asn Phe Thr Ile Thr Asn Leu Arg 13790 13795 13800 Tyr Glu Glu Asn Met Trp Pro Gly Ser Arg Lys Phe Asn Thr Thr 13805 13810 13815 Glu Arg Val Leu Gln Gly Leu Leu Arg Pro Leu Phe Lys Asn Thr 13820 13825 13830 Ser Val Gly Pro Leu Tyr Ser Gly Cys Arg Leu Thr Leu Leu Arg 13835 13840 13845 Pro Glu Lys Asp Gly Glu Ala Thr Gly Val Asp Ala Ile Cys Thr 13850 13855 13860 His Arg Pro Asp Pro Thr Gly Pro Gly Leu Asp Arg Glu Gln Leu 13865 13870 13875 Tyr Leu Glu Leu Ser Gln Leu Thr His Ser Ile Thr Glu Leu Gly 13880 13885 13890 Pro Tyr Thr Leu Asp Arg Asp Ser Leu Tyr Val Asn Gly Phe Thr 13895 13900 13905 His Arg Ser Ser Val Pro Thr Thr Ser Thr Gly Val Val Ser Glu 13910 13915 13920 Glu Pro Phe Thr Leu Asn Phe Thr Ile Asn Asn Leu Arg Tyr Met 13925 13930 13935 Ala Asp Met Gly Gln Pro Gly Ser Leu Lys Phe Asn Ile Thr Asp 13940 13945 13950 Asn Val Met Gln His Leu Leu Ser Pro Leu Phe Gln Arg Ser Ser 13955 13960 13965 Leu Gly Ala Arg Tyr Thr Gly Cys Arg Val Ile Ala Leu Arg Ser 13970 13975 13980 Val Lys Asn Gly Ala Glu Thr Arg Val Asp Leu Leu Cys Thr Tyr 13985 13990 13995 Leu Gln Pro Leu Ser Gly Pro Gly Leu Pro Ile Lys Gln Val Phe 14000 14005 14010 His Glu Leu Ser Gln Gln Thr His Gly Ile Thr Arg Leu Gly Pro 14015 14020 14025 Tyr Ser Leu Asp Lys Asp Ser Leu Tyr Leu Asn Gly Tyr Asn Glu 14030 14035 14040 Pro Gly Pro Asp Glu Pro Pro Thr Thr Pro Lys Pro Ala Thr Thr 14045 14050 14055 Phe Leu Pro Pro Leu Ser Glu Ala Thr Thr Ala Met Gly Tyr His 14060 14065 14070 Leu Lys Thr Leu Thr Leu Asn Phe Thr Ile Ser Asn Leu Gln Tyr 14075 14080 14085 Ser Pro Asp Met Gly Lys Gly Ser Ala Thr Phe Asn Ser Thr Glu 14090 14095 14100 Gly Val Leu Gln His Leu Leu Arg Pro Leu Phe Gln Lys Ser Ser 14105 14110 14115 Met Gly Pro Phe Tyr Leu Gly Cys Gln Leu Ile Ser Leu Arg Pro 14120 14125 14130 Glu Lys Asp Gly Ala Ala Thr Gly Val Asp Thr Thr Cys Thr Tyr 14135 14140 14145 His Pro Asp Pro Val Gly Pro Gly Leu Asp Ile Gln Gln Leu Tyr 14150 14155 14160 Trp Glu Leu Ser Gln Leu Thr His Gly Val Thr Gln Leu Gly Phe 14165 14170 14175 Tyr Val Leu Asp Arg Asp Ser Leu Phe Ile Asn Gly Tyr Ala Pro 14180 14185 14190 Gln Asn Leu Ser Ile Arg Gly Glu Tyr Gln Ile Asn Phe His Ile 14195 14200 14205 Val Asn Trp Asn Leu Ser Asn Pro Asp Pro Thr Ser Ser Glu Tyr 14210 14215 14220 Ile Thr Leu Leu Arg Asp Ile Gln Asp Lys Val Thr Thr Leu Tyr 14225 14230 14235 Lys Gly Ser Gln Leu His Asp Thr Phe Arg Phe Cys Leu Val Thr 14240 14245 14250 Asn Leu Thr Met Asp Ser Val Leu Val Thr Val Lys Ala Leu Phe 14255 14260 14265 Ser Ser Asn Leu Asp Pro Ser Leu Val Glu Gln Val Phe Leu Asp 14270 14275 14280 Lys Thr Leu Asn Ala Ser Phe His Trp Leu Gly Ser Thr Tyr Gln 14285 14290 14295 Leu Val Asp Ile His Val Thr Glu Met Glu Ser Ser Val Tyr Gln 14300 14305 14310 Pro Thr Ser Ser Ser Ser Thr Gln His Phe Tyr Leu Asn Phe Thr 14315 14320 14325 Ile Thr Asn Leu Pro Tyr Ser Gln Asp Lys Ala Gln Pro Gly Thr 14330 14335 14340 Thr Asn Tyr Gln Arg Asn Lys Arg Asn Ile Glu Asp Ala Leu Asn 14345 14350 14355 Gln Leu Phe Arg Asn Ser Ser Ile Lys Ser Tyr Phe Ser Asp Cys 14360 14365 14370 Gln Val Ser Thr Phe Arg Ser Val Pro Asn Arg His His Thr Gly 14375 14380 14385 Val Asp Ser Leu Cys Asn Phe Ser Pro Leu Ala Arg Arg Val Asp 14390 14395 14400 Arg Val Ala Ile Tyr Glu Glu Phe Leu Arg Met Thr Arg Asn Gly 14405 14410 14415 Thr Gln Leu Gln Asn Phe Thr Leu Asp Arg Ser Ser Val Leu Val 14420 14425 14430 Asp Gly Tyr Ser Pro Asn Arg Asn Glu Pro Leu Thr Gly Asn Ser 14435 14440 14445 Asp Leu Pro Phe Trp Ala Val Ile Leu Ile Gly Leu Ala Gly Leu 14450 14455 14460 Leu Gly Val Ile Thr Cys Leu Ile Cys Gly Val Leu Val Thr Thr 14465 14470 14475 Arg Arg Arg Lys Lys Glu Gly Glu Tyr Asn Val Gln Gln Gln Cys 14480 14485 14490 Pro Gly Tyr Tyr Gln Ser His Leu Asp Leu Glu Asp Leu Gln 14495 14500 14505 58630PRTHomo sapiens 58Met Ala Leu Pro Thr Ala Arg Pro Leu Leu Gly Ser Cys Gly Thr Pro 1 5 10 15 Ala Leu Gly Ser Leu Leu Phe Leu Leu Phe Ser Leu Gly Trp Val Gln 20 25 30 Pro Ser Arg Thr Leu Ala Gly Glu Thr Gly Gln Glu Ala Ala Pro Leu 35 40 45 Asp Gly Val Leu Ala Asn Pro Pro Asn Ile Ser Ser Leu Ser Pro Arg 50 55 60 Gln Leu Leu Gly Phe Pro Cys Ala Glu Val Ser Gly Leu Ser Thr Glu 65 70 75 80 Arg Val Arg Glu Leu Ala Val Ala Leu Ala Gln Lys Asn Val Lys Leu 85 90 95 Ser Thr Glu Gln Leu Arg Cys Leu Ala His Arg Leu Ser Glu Pro Pro 100 105 110 Glu Asp Leu Asp Ala Leu Pro Leu Asp Leu Leu Leu Phe Leu Asn Pro 115 120 125 Asp Ala Phe Ser Gly Pro Gln Ala Cys Thr Arg Phe Phe Ser Arg Ile 130 135 140 Thr Lys Ala Asn Val Asp Leu Leu Pro Arg Gly Ala Pro Glu Arg Gln 145 150 155 160 Arg Leu Leu Pro Ala Ala Leu Ala Cys Trp Gly Val Arg Gly Ser Leu 165 170 175 Leu Ser Glu Ala Asp Val Arg Ala Leu Gly Gly Leu Ala Cys Asp Leu 180 185 190 Pro Gly Arg Phe Val Ala Glu Ser Ala Glu Val Leu Leu Pro Arg Leu 195 200 205 Val Ser Cys Pro Gly Pro Leu Asp Gln Asp Gln Gln Glu Ala Ala Arg 210 215 220 Ala Ala Leu Gln Gly Gly Gly Pro Pro Tyr Gly Pro Pro Ser Thr Trp 225 230 235 240 Ser Val Ser Thr Met Asp Ala Leu Arg Gly Leu Leu Pro Val Leu Gly 245 250 255 Gln Pro Ile Ile Arg Ser Ile Pro Gln Gly Ile Val Ala Ala Trp Arg 260 265 270 Gln Arg Ser Ser Arg Asp Pro Ser Trp Arg Gln Pro Glu Arg Thr Ile 275 280 285 Leu Arg Pro Arg Phe Arg Arg Glu Val Glu Lys Thr Ala Cys Pro Ser 290 295 300 Gly Lys Lys Ala Arg Glu Ile Asp Glu Ser Leu Ile Phe Tyr Lys Lys 305 310 315 320 Trp Glu Leu Glu Ala Cys Val Asp Ala Ala Leu

Leu Ala Thr Gln Met 325 330 335 Asp Arg Val Asn Ala Ile Pro Phe Thr Tyr Glu Gln Leu Asp Val Leu 340 345 350 Lys His Lys Leu Asp Glu Leu Tyr Pro Gln Gly Tyr Pro Glu Ser Val 355 360 365 Ile Gln His Leu Gly Tyr Leu Phe Leu Lys Met Ser Pro Glu Asp Ile 370 375 380 Arg Lys Trp Asn Val Thr Ser Leu Glu Thr Leu Lys Ala Leu Leu Glu 385 390 395 400 Val Asn Lys Gly His Glu Met Ser Pro Gln Ala Pro Arg Arg Pro Leu 405 410 415 Pro Gln Val Ala Thr Leu Ile Asp Arg Phe Val Lys Gly Arg Gly Gln 420 425 430 Leu Asp Lys Asp Thr Leu Asp Thr Leu Thr Ala Phe Tyr Pro Gly Tyr 435 440 445 Leu Cys Ser Leu Ser Pro Glu Glu Leu Ser Ser Val Pro Pro Ser Ser 450 455 460 Ile Trp Ala Val Arg Pro Gln Asp Leu Asp Thr Cys Asp Pro Arg Gln 465 470 475 480 Leu Asp Val Leu Tyr Pro Lys Ala Arg Leu Ala Phe Gln Asn Met Asn 485 490 495 Gly Ser Glu Tyr Phe Val Lys Ile Gln Ser Phe Leu Gly Gly Ala Pro 500 505 510 Thr Glu Asp Leu Lys Ala Leu Ser Gln Gln Asn Val Ser Met Asp Leu 515 520 525 Ala Thr Phe Met Lys Leu Arg Thr Asp Ala Val Leu Pro Leu Thr Val 530 535 540 Ala Glu Val Gln Lys Leu Leu Gly Pro His Val Glu Gly Leu Lys Ala 545 550 555 560 Glu Glu Arg His Arg Pro Val Arg Asp Trp Ile Leu Arg Gln Arg Gln 565 570 575 Asp Asp Leu Asp Thr Leu Gly Leu Gly Leu Gln Gly Gly Ile Pro Asn 580 585 590 Gly Tyr Leu Val Leu Asp Leu Ser Met Gln Glu Ala Leu Ser Gly Thr 595 600 605 Pro Cys Leu Leu Gly Pro Gly Pro Val Leu Thr Val Leu Ala Leu Leu 610 615 620 Leu Ala Ser Thr Leu Ala 625 630 59622PRTHomo sapiens 59Met Ala Leu Pro Thr Ala Arg Pro Leu Leu Gly Ser Cys Gly Thr Pro 1 5 10 15 Ala Leu Gly Ser Leu Leu Phe Leu Leu Phe Ser Leu Gly Trp Val Gln 20 25 30 Pro Ser Arg Thr Leu Ala Gly Glu Thr Gly Gln Glu Ala Ala Pro Leu 35 40 45 Asp Gly Val Leu Ala Asn Pro Pro Asn Ile Ser Ser Leu Ser Pro Arg 50 55 60 Gln Leu Leu Gly Phe Pro Cys Ala Glu Val Ser Gly Leu Ser Thr Glu 65 70 75 80 Arg Val Arg Glu Leu Ala Val Ala Leu Ala Gln Lys Asn Val Lys Leu 85 90 95 Ser Thr Glu Gln Leu Arg Cys Leu Ala His Arg Leu Ser Glu Pro Pro 100 105 110 Glu Asp Leu Asp Ala Leu Pro Leu Asp Leu Leu Leu Phe Leu Asn Pro 115 120 125 Asp Ala Phe Ser Gly Pro Gln Ala Cys Thr Arg Phe Phe Ser Arg Ile 130 135 140 Thr Lys Ala Asn Val Asp Leu Leu Pro Arg Gly Ala Pro Glu Arg Gln 145 150 155 160 Arg Leu Leu Pro Ala Ala Leu Ala Cys Trp Gly Val Arg Gly Ser Leu 165 170 175 Leu Ser Glu Ala Asp Val Arg Ala Leu Gly Gly Leu Ala Cys Asp Leu 180 185 190 Pro Gly Arg Phe Val Ala Glu Ser Ala Glu Val Leu Leu Pro Arg Leu 195 200 205 Val Ser Cys Pro Gly Pro Leu Asp Gln Asp Gln Gln Glu Ala Ala Arg 210 215 220 Ala Ala Leu Gln Gly Gly Gly Pro Pro Tyr Gly Pro Pro Ser Thr Trp 225 230 235 240 Ser Val Ser Thr Met Asp Ala Leu Arg Gly Leu Leu Pro Val Leu Gly 245 250 255 Gln Pro Ile Ile Arg Ser Ile Pro Gln Gly Ile Val Ala Ala Trp Arg 260 265 270 Gln Arg Ser Ser Arg Asp Pro Ser Trp Arg Gln Pro Glu Arg Thr Ile 275 280 285 Leu Arg Pro Arg Phe Arg Arg Glu Val Glu Lys Thr Ala Cys Pro Ser 290 295 300 Gly Lys Lys Ala Arg Glu Ile Asp Glu Ser Leu Ile Phe Tyr Lys Lys 305 310 315 320 Trp Glu Leu Glu Ala Cys Val Asp Ala Ala Leu Leu Ala Thr Gln Met 325 330 335 Asp Arg Val Asn Ala Ile Pro Phe Thr Tyr Glu Gln Leu Asp Val Leu 340 345 350 Lys His Lys Leu Asp Glu Leu Tyr Pro Gln Gly Tyr Pro Glu Ser Val 355 360 365 Ile Gln His Leu Gly Tyr Leu Phe Leu Lys Met Ser Pro Glu Asp Ile 370 375 380 Arg Lys Trp Asn Val Thr Ser Leu Glu Thr Leu Lys Ala Leu Leu Glu 385 390 395 400 Val Asn Lys Gly His Glu Met Ser Pro Gln Val Ala Thr Leu Ile Asp 405 410 415 Arg Phe Val Lys Gly Arg Gly Gln Leu Asp Lys Asp Thr Leu Asp Thr 420 425 430 Leu Thr Ala Phe Tyr Pro Gly Tyr Leu Cys Ser Leu Ser Pro Glu Glu 435 440 445 Leu Ser Ser Val Pro Pro Ser Ser Ile Trp Ala Val Arg Pro Gln Asp 450 455 460 Leu Asp Thr Cys Asp Pro Arg Gln Leu Asp Val Leu Tyr Pro Lys Ala 465 470 475 480 Arg Leu Ala Phe Gln Asn Met Asn Gly Ser Glu Tyr Phe Val Lys Ile 485 490 495 Gln Ser Phe Leu Gly Gly Ala Pro Thr Glu Asp Leu Lys Ala Leu Ser 500 505 510 Gln Gln Asn Val Ser Met Asp Leu Ala Thr Phe Met Lys Leu Arg Thr 515 520 525 Asp Ala Val Leu Pro Leu Thr Val Ala Glu Val Gln Lys Leu Leu Gly 530 535 540 Pro His Val Glu Gly Leu Lys Ala Glu Glu Arg His Arg Pro Val Arg 545 550 555 560 Asp Trp Ile Leu Arg Gln Arg Gln Asp Asp Leu Asp Thr Leu Gly Leu 565 570 575 Gly Leu Gln Gly Gly Ile Pro Asn Gly Tyr Leu Val Leu Asp Leu Ser 580 585 590 Met Gln Glu Ala Leu Ser Gly Thr Pro Cys Leu Leu Gly Pro Gly Pro 595 600 605 Val Leu Thr Val Leu Ala Leu Leu Leu Ala Ser Thr Leu Ala 610 615 620 60300PRTHomo sapiens 60Met Arg Ile Ala Val Ile Cys Phe Cys Leu Leu Gly Ile Thr Cys Ala 1 5 10 15 Ile Pro Val Lys Gln Ala Asp Ser Gly Ser Ser Glu Glu Lys Gln Leu 20 25 30 Tyr Asn Lys Tyr Pro Asp Ala Val Ala Thr Trp Leu Asn Pro Asp Pro 35 40 45 Ser Gln Lys Gln Asn Leu Leu Ala Pro Gln Thr Leu Pro Ser Lys Ser 50 55 60 Asn Glu Ser His Asp His Met Asp Asp Met Asp Asp Glu Asp Asp Asp 65 70 75 80 Asp His Val Asp Ser Gln Asp Ser Ile Asp Ser Asn Asp Ser Asp Asp 85 90 95 Val Asp Asp Thr Asp Asp Ser His Gln Ser Asp Glu Ser His His Ser 100 105 110 Asp Glu Ser Asp Glu Leu Val Thr Asp Phe Pro Thr Asp Leu Pro Ala 115 120 125 Thr Glu Val Phe Thr Pro Val Val Pro Thr Val Asp Thr Tyr Asp Gly 130 135 140 Arg Gly Asp Ser Val Val Tyr Gly Leu Arg Ser Lys Ser Lys Lys Phe 145 150 155 160 Arg Arg Pro Asp Ile Gln Tyr Pro Asp Ala Thr Asp Glu Asp Ile Thr 165 170 175 Ser His Met Glu Ser Glu Glu Leu Asn Gly Ala Tyr Lys Ala Ile Pro 180 185 190 Val Ala Gln Asp Leu Asn Ala Pro Ser Asp Trp Asp Ser Arg Gly Lys 195 200 205 Asp Ser Tyr Glu Thr Ser Gln Leu Asp Asp Gln Ser Ala Glu Thr His 210 215 220 Ser His Lys Gln Ser Arg Leu Tyr Lys Arg Lys Ala Asn Asp Glu Ser 225 230 235 240 Asn Glu His Ser Asp Val Ile Asp Ser Gln Glu Leu Ser Lys Val Ser 245 250 255 Arg Glu Phe His Ser His Glu Phe His Ser His Glu Asp Met Leu Val 260 265 270 Val Asp Pro Lys Ser Lys Glu Glu Asp Lys His Leu Lys Phe Arg Ile 275 280 285 Ser His Glu Leu Asp Ser Ala Ser Ser Glu Val Asn 290 295 300 61314PRTHomo sapiens 61Met Arg Ile Ala Val Ile Cys Phe Cys Leu Leu Gly Ile Thr Cys Ala 1 5 10 15 Ile Pro Val Lys Gln Ala Asp Ser Gly Ser Ser Glu Glu Lys Gln Leu 20 25 30 Tyr Asn Lys Tyr Pro Asp Ala Val Ala Thr Trp Leu Asn Pro Asp Pro 35 40 45 Ser Gln Lys Gln Asn Leu Leu Ala Pro Gln Asn Ala Val Ser Ser Glu 50 55 60 Glu Thr Asn Asp Phe Lys Gln Glu Thr Leu Pro Ser Lys Ser Asn Glu 65 70 75 80 Ser His Asp His Met Asp Asp Met Asp Asp Glu Asp Asp Asp Asp His 85 90 95 Val Asp Ser Gln Asp Ser Ile Asp Ser Asn Asp Ser Asp Asp Val Asp 100 105 110 Asp Thr Asp Asp Ser His Gln Ser Asp Glu Ser His His Ser Asp Glu 115 120 125 Ser Asp Glu Leu Val Thr Asp Phe Pro Thr Asp Leu Pro Ala Thr Glu 130 135 140 Val Phe Thr Pro Val Val Pro Thr Val Asp Thr Tyr Asp Gly Arg Gly 145 150 155 160 Asp Ser Val Val Tyr Gly Leu Arg Ser Lys Ser Lys Lys Phe Arg Arg 165 170 175 Pro Asp Ile Gln Tyr Pro Asp Ala Thr Asp Glu Asp Ile Thr Ser His 180 185 190 Met Glu Ser Glu Glu Leu Asn Gly Ala Tyr Lys Ala Ile Pro Val Ala 195 200 205 Gln Asp Leu Asn Ala Pro Ser Asp Trp Asp Ser Arg Gly Lys Asp Ser 210 215 220 Tyr Glu Thr Ser Gln Leu Asp Asp Gln Ser Ala Glu Thr His Ser His 225 230 235 240 Lys Gln Ser Arg Leu Tyr Lys Arg Lys Ala Asn Asp Glu Ser Asn Glu 245 250 255 His Ser Asp Val Ile Asp Ser Gln Glu Leu Ser Lys Val Ser Arg Glu 260 265 270 Phe His Ser His Glu Phe His Ser His Glu Asp Met Leu Val Val Asp 275 280 285 Pro Lys Ser Lys Glu Glu Asp Lys His Leu Lys Phe Arg Ile Ser His 290 295 300 Glu Leu Asp Ser Ala Ser Ser Glu Val Asn 305 310 62287PRTHomo sapiens 62Met Arg Ile Ala Val Ile Cys Phe Cys Leu Leu Gly Ile Thr Cys Ala 1 5 10 15 Ile Pro Val Lys Gln Ala Asp Ser Gly Ser Ser Glu Glu Lys Gln Asn 20 25 30 Ala Val Ser Ser Glu Glu Thr Asn Asp Phe Lys Gln Glu Thr Leu Pro 35 40 45 Ser Lys Ser Asn Glu Ser His Asp His Met Asp Asp Met Asp Asp Glu 50 55 60 Asp Asp Asp Asp His Val Asp Ser Gln Asp Ser Ile Asp Ser Asn Asp 65 70 75 80 Ser Asp Asp Val Asp Asp Thr Asp Asp Ser His Gln Ser Asp Glu Ser 85 90 95 His His Ser Asp Glu Ser Asp Glu Leu Val Thr Asp Phe Pro Thr Asp 100 105 110 Leu Pro Ala Thr Glu Val Phe Thr Pro Val Val Pro Thr Val Asp Thr 115 120 125 Tyr Asp Gly Arg Gly Asp Ser Val Val Tyr Gly Leu Arg Ser Lys Ser 130 135 140 Lys Lys Phe Arg Arg Pro Asp Ile Gln Tyr Pro Asp Ala Thr Asp Glu 145 150 155 160 Asp Ile Thr Ser His Met Glu Ser Glu Glu Leu Asn Gly Ala Tyr Lys 165 170 175 Ala Ile Pro Val Ala Gln Asp Leu Asn Ala Pro Ser Asp Trp Asp Ser 180 185 190 Arg Gly Lys Asp Ser Tyr Glu Thr Ser Gln Leu Asp Asp Gln Ser Ala 195 200 205 Glu Thr His Ser His Lys Gln Ser Arg Leu Tyr Lys Arg Lys Ala Asn 210 215 220 Asp Glu Ser Asn Glu His Ser Asp Val Ile Asp Ser Gln Glu Leu Ser 225 230 235 240 Lys Val Ser Arg Glu Phe His Ser His Glu Phe His Ser His Glu Asp 245 250 255 Met Leu Val Val Asp Pro Lys Ser Lys Glu Glu Asp Lys His Leu Lys 260 265 270 Phe Arg Ile Ser His Glu Leu Asp Ser Ala Ser Ser Glu Val Asn 275 280 285 63411PRTHomo sapiens 63Met Val Cys Phe Arg Leu Phe Pro Val Pro Gly Ser Gly Leu Val Leu 1 5 10 15 Val Cys Leu Val Leu Gly Ala Val Arg Ser Tyr Ala Leu Glu Leu Asn 20 25 30 Leu Thr Asp Ser Glu Asn Ala Thr Cys Leu Tyr Ala Lys Trp Gln Met 35 40 45 Asn Phe Thr Val Arg Tyr Glu Thr Thr Asn Lys Thr Tyr Lys Thr Val 50 55 60 Thr Ile Ser Asp His Gly Thr Val Thr Tyr Asn Gly Ser Ile Cys Gly 65 70 75 80 Asp Asp Gln Asn Gly Pro Lys Ile Ala Val Gln Phe Gly Pro Gly Phe 85 90 95 Ser Trp Ile Ala Asn Phe Thr Lys Ala Ala Ser Thr Tyr Ser Ile Asp 100 105 110 Ser Val Ser Phe Ser Tyr Asn Thr Gly Asp Asn Thr Thr Phe Pro Asp 115 120 125 Ala Glu Asp Lys Gly Ile Leu Thr Val Asp Glu Leu Leu Ala Ile Arg 130 135 140 Ile Pro Leu Asn Asp Leu Phe Arg Cys Asn Ser Leu Ser Thr Leu Glu 145 150 155 160 Lys Asn Asp Val Val Gln His Tyr Trp Asp Val Leu Val Gln Ala Phe 165 170 175 Val Gln Asn Gly Thr Val Ser Thr Asn Glu Phe Leu Cys Asp Lys Asp 180 185 190 Lys Thr Ser Thr Val Ala Pro Thr Ile His Thr Thr Val Pro Ser Pro 195 200 205 Thr Thr Thr Pro Thr Pro Lys Glu Lys Pro Glu Ala Gly Thr Tyr Ser 210 215 220 Val Asn Asn Gly Asn Asp Thr Cys Leu Leu Ala Thr Met Gly Leu Gln 225 230 235 240 Leu Asn Ile Thr Gln Asp Lys Val Ala Ser Val Ile Asn Ile Asn Pro 245 250 255 Asn Thr Thr His Ser Thr Gly Ser Cys Arg Ser His Thr Ala Leu Leu 260 265 270 Arg Leu Asn Ser Ser Thr Ile Lys Tyr Leu Asp Phe Val Phe Ala Val 275 280 285 Lys Asn Glu Asn Arg Phe Tyr Leu Lys Glu Val Asn Ile Ser Met Tyr 290 295 300 Leu Val Asn Gly Ser Val Phe Ser Ile Ala Asn Asn Asn Leu Ser Tyr 305 310 315 320 Trp Asp Ala Pro Leu Gly Ser Ser Tyr Met Cys Asn Lys Glu Gln Thr 325 330 335 Val Ser Val Ser Gly Ala Phe Gln Ile Asn Thr Phe Asp Leu Arg Val 340 345 350 Gln Pro Phe Asn Val Thr Gln Gly Lys Tyr Ser Thr Ala Glu Glu Cys 355 360 365 Ser Ala Asp Ser Asp Leu Asn Phe Leu Ile Pro Val Ala Val Gly Val 370 375 380 Ala Leu Gly Phe Leu Ile Ile Val Val Phe Ile Ser Tyr Met Ile Gly 385 390 395 400 Arg Arg Lys Ser Arg Thr Gly Tyr Gln Ser Val 405 410 64410PRTHomo sapiens 64Met Val Cys Phe Arg Leu Phe Pro Val Pro Gly Ser Gly Leu Val Leu 1 5 10 15 Val Cys Leu Val Leu Gly Ala Val Arg Ser Tyr Ala Leu Glu Leu Asn 20 25 30 Leu Thr Asp Ser Glu Asn Ala Thr Cys Leu Tyr Ala Lys Trp Gln Met 35 40 45 Asn Phe Thr Val Arg Tyr Glu Thr

Thr Asn Lys Thr Tyr Lys Thr Val 50 55 60 Thr Ile Ser Asp His Gly Thr Val Thr Tyr Asn Gly Ser Ile Cys Gly 65 70 75 80 Asp Asp Gln Asn Gly Pro Lys Ile Ala Val Gln Phe Gly Pro Gly Phe 85 90 95 Ser Trp Ile Ala Asn Phe Thr Lys Ala Ala Ser Thr Tyr Ser Ile Asp 100 105 110 Ser Val Ser Phe Ser Tyr Asn Thr Gly Asp Asn Thr Thr Phe Pro Asp 115 120 125 Ala Glu Asp Lys Gly Ile Leu Thr Val Asp Glu Leu Leu Ala Ile Arg 130 135 140 Ile Pro Leu Asn Asp Leu Phe Arg Cys Asn Ser Leu Ser Thr Leu Glu 145 150 155 160 Lys Asn Asp Val Val Gln His Tyr Trp Asp Val Leu Val Gln Ala Phe 165 170 175 Val Gln Asn Gly Thr Val Ser Thr Asn Glu Phe Leu Cys Asp Lys Asp 180 185 190 Lys Thr Ser Thr Val Ala Pro Thr Ile His Thr Thr Val Pro Ser Pro 195 200 205 Thr Thr Thr Pro Thr Pro Lys Glu Lys Pro Glu Ala Gly Thr Tyr Ser 210 215 220 Val Asn Asn Gly Asn Asp Thr Cys Leu Leu Ala Thr Met Gly Leu Gln 225 230 235 240 Leu Asn Ile Thr Gln Asp Lys Val Ala Ser Val Ile Asn Ile Asn Pro 245 250 255 Asn Thr Thr His Ser Thr Gly Ser Cys Arg Ser His Thr Ala Leu Leu 260 265 270 Arg Leu Asn Ser Ser Thr Ile Lys Tyr Leu Asp Phe Val Phe Ala Val 275 280 285 Lys Asn Glu Asn Arg Phe Tyr Leu Lys Glu Val Asn Ile Ser Met Tyr 290 295 300 Leu Val Asn Gly Ser Val Phe Ser Ile Ala Asn Asn Asn Leu Ser Tyr 305 310 315 320 Trp Asp Ala Pro Leu Gly Ser Ser Tyr Met Cys Asn Lys Glu Gln Thr 325 330 335 Val Ser Val Ser Gly Ala Phe Gln Ile Asn Thr Phe Asp Leu Arg Val 340 345 350 Gln Pro Phe Asn Val Thr Gln Gly Lys Tyr Ser Thr Ala Gln Asp Cys 355 360 365 Ser Ala Asp Asp Asp Asn Phe Leu Val Pro Ile Ala Val Gly Ala Ala 370 375 380 Leu Ala Gly Val Leu Ile Leu Val Leu Leu Ala Tyr Phe Ile Gly Leu 385 390 395 400 Lys His His His Ala Gly Tyr Glu Gln Phe 405 410 65410PRTHomo sapiens 65Met Val Cys Phe Arg Leu Phe Pro Val Pro Gly Ser Gly Leu Val Leu 1 5 10 15 Val Cys Leu Val Leu Gly Ala Val Arg Ser Tyr Ala Leu Glu Leu Asn 20 25 30 Leu Thr Asp Ser Glu Asn Ala Thr Cys Leu Tyr Ala Lys Trp Gln Met 35 40 45 Asn Phe Thr Val Arg Tyr Glu Thr Thr Asn Lys Thr Tyr Lys Thr Val 50 55 60 Thr Ile Ser Asp His Gly Thr Val Thr Tyr Asn Gly Ser Ile Cys Gly 65 70 75 80 Asp Asp Gln Asn Gly Pro Lys Ile Ala Val Gln Phe Gly Pro Gly Phe 85 90 95 Ser Trp Ile Ala Asn Phe Thr Lys Ala Ala Ser Thr Tyr Ser Ile Asp 100 105 110 Ser Val Ser Phe Ser Tyr Asn Thr Gly Asp Asn Thr Thr Phe Pro Asp 115 120 125 Ala Glu Asp Lys Gly Ile Leu Thr Val Asp Glu Leu Leu Ala Ile Arg 130 135 140 Ile Pro Leu Asn Asp Leu Phe Arg Cys Asn Ser Leu Ser Thr Leu Glu 145 150 155 160 Lys Asn Asp Val Val Gln His Tyr Trp Asp Val Leu Val Gln Ala Phe 165 170 175 Val Gln Asn Gly Thr Val Ser Thr Asn Glu Phe Leu Cys Asp Lys Asp 180 185 190 Lys Thr Ser Thr Val Ala Pro Thr Ile His Thr Thr Val Pro Ser Pro 195 200 205 Thr Thr Thr Pro Thr Pro Lys Glu Lys Pro Glu Ala Gly Thr Tyr Ser 210 215 220 Val Asn Asn Gly Asn Asp Thr Cys Leu Leu Ala Thr Met Gly Leu Gln 225 230 235 240 Leu Asn Ile Thr Gln Asp Lys Val Ala Ser Val Ile Asn Ile Asn Pro 245 250 255 Asn Thr Thr His Ser Thr Gly Ser Cys Arg Ser His Thr Ala Leu Leu 260 265 270 Arg Leu Asn Ser Ser Thr Ile Lys Tyr Leu Asp Phe Val Phe Ala Val 275 280 285 Lys Asn Glu Asn Arg Phe Tyr Leu Lys Glu Val Asn Ile Ser Met Tyr 290 295 300 Leu Val Asn Gly Ser Val Phe Ser Ile Ala Asn Asn Asn Leu Ser Tyr 305 310 315 320 Trp Asp Ala Pro Leu Gly Ser Ser Tyr Met Cys Asn Lys Glu Gln Thr 325 330 335 Val Ser Val Ser Gly Ala Phe Gln Ile Asn Thr Phe Asp Leu Arg Val 340 345 350 Gln Pro Phe Asn Val Thr Gln Gly Lys Tyr Ser Thr Ala Gln Glu Cys 355 360 365 Ser Leu Asp Asp Asp Thr Ile Leu Ile Pro Ile Ile Val Gly Ala Gly 370 375 380 Leu Ser Gly Leu Ile Ile Val Ile Val Ile Ala Tyr Val Ile Gly Arg 385 390 395 400 Arg Lys Ser Tyr Ala Gly Tyr Gln Thr Leu 405 410 66262PRTHomo sapiens 66Met Trp Phe Leu Val Leu Cys Leu Ala Leu Ser Leu Gly Gly Thr Gly 1 5 10 15 Ala Ala Pro Pro Ile Gln Ser Arg Ile Val Gly Gly Trp Glu Cys Glu 20 25 30 Gln His Ser Gln Pro Trp Gln Ala Ala Leu Tyr His Phe Ser Thr Phe 35 40 45 Gln Cys Gly Gly Ile Leu Val His Arg Gln Trp Val Leu Thr Ala Ala 50 55 60 His Cys Ile Ser Asp Asn Tyr Gln Leu Trp Leu Gly Arg His Asn Leu 65 70 75 80 Phe Asp Asp Glu Asn Thr Ala Gln Phe Val His Val Ser Glu Ser Phe 85 90 95 Pro His Pro Gly Phe Asn Met Ser Leu Leu Glu Asn His Thr Arg Gln 100 105 110 Ala Asp Glu Asp Tyr Ser His Asp Leu Met Leu Leu Arg Leu Thr Glu 115 120 125 Pro Ala Asp Thr Ile Thr Asp Ala Val Lys Val Val Glu Leu Pro Thr 130 135 140 Glu Glu Pro Glu Val Gly Ser Thr Cys Leu Ala Ser Gly Trp Gly Ser 145 150 155 160 Ile Glu Pro Glu Asn Phe Ser Phe Pro Asp Asp Leu Gln Cys Val Asp 165 170 175 Leu Lys Ile Leu Pro Asn Asp Glu Cys Lys Lys Ala His Val Gln Lys 180 185 190 Val Thr Asp Phe Met Leu Cys Val Gly His Leu Glu Gly Gly Lys Asp 195 200 205 Thr Cys Val Gly Asp Ser Gly Gly Pro Leu Met Cys Asp Gly Val Leu 210 215 220 Gln Gly Val Thr Ser Trp Gly Tyr Val Pro Cys Gly Thr Pro Asn Lys 225 230 235 240 Pro Ser Val Ala Val Arg Val Leu Ser Tyr Val Lys Trp Ile Glu Asp 245 250 255 Thr Ile Ala Glu Asn Ser 260 67860PRTHomo sapiens 67Met Gly Pro Trp Gly Trp Lys Leu Arg Trp Thr Val Ala Leu Leu Leu 1 5 10 15 Ala Ala Ala Gly Thr Ala Val Gly Asp Arg Cys Glu Arg Asn Glu Phe 20 25 30 Gln Cys Gln Asp Gly Lys Cys Ile Ser Tyr Lys Trp Val Cys Asp Gly 35 40 45 Ser Ala Glu Cys Gln Asp Gly Ser Asp Glu Ser Gln Glu Thr Cys Leu 50 55 60 Ser Val Thr Cys Lys Ser Gly Asp Phe Ser Cys Gly Gly Arg Val Asn 65 70 75 80 Arg Cys Ile Pro Gln Phe Trp Arg Cys Asp Gly Gln Val Asp Cys Asp 85 90 95 Asn Gly Ser Asp Glu Gln Gly Cys Pro Pro Lys Thr Cys Ser Gln Asp 100 105 110 Glu Phe Arg Cys His Asp Gly Lys Cys Ile Ser Arg Gln Phe Val Cys 115 120 125 Asp Ser Asp Arg Asp Cys Leu Asp Gly Ser Asp Glu Ala Ser Cys Pro 130 135 140 Val Leu Thr Cys Gly Pro Ala Ser Phe Gln Cys Asn Ser Ser Thr Cys 145 150 155 160 Ile Pro Gln Leu Trp Ala Cys Asp Asn Asp Pro Asp Cys Glu Asp Gly 165 170 175 Ser Asp Glu Trp Pro Gln Arg Cys Arg Gly Leu Tyr Val Phe Gln Gly 180 185 190 Asp Ser Ser Pro Cys Ser Ala Phe Glu Phe His Cys Leu Ser Gly Glu 195 200 205 Cys Ile His Ser Ser Trp Arg Cys Asp Gly Gly Pro Asp Cys Lys Asp 210 215 220 Lys Ser Asp Glu Glu Asn Cys Ala Val Ala Thr Cys Arg Pro Asp Glu 225 230 235 240 Phe Gln Cys Ser Asp Gly Asn Cys Ile His Gly Ser Arg Gln Cys Asp 245 250 255 Arg Glu Tyr Asp Cys Lys Asp Met Ser Asp Glu Val Gly Cys Val Asn 260 265 270 Val Thr Leu Cys Glu Gly Pro Asn Lys Phe Lys Cys His Ser Gly Glu 275 280 285 Cys Ile Thr Leu Asp Lys Val Cys Asn Met Ala Arg Asp Cys Arg Asp 290 295 300 Trp Ser Asp Glu Pro Ile Lys Glu Cys Gly Thr Asn Glu Cys Leu Asp 305 310 315 320 Asn Asn Gly Gly Cys Ser His Val Cys Asn Asp Leu Lys Ile Gly Tyr 325 330 335 Glu Cys Leu Cys Pro Asp Gly Phe Gln Leu Val Ala Gln Arg Arg Cys 340 345 350 Glu Asp Ile Asp Glu Cys Gln Asp Pro Asp Thr Cys Ser Gln Leu Cys 355 360 365 Val Asn Leu Glu Gly Gly Tyr Lys Cys Gln Cys Glu Glu Gly Phe Gln 370 375 380 Leu Asp Pro His Thr Lys Ala Cys Lys Ala Val Gly Ser Ile Ala Tyr 385 390 395 400 Leu Phe Phe Thr Asn Arg His Glu Val Arg Lys Met Thr Leu Asp Arg 405 410 415 Ser Glu Tyr Thr Ser Leu Ile Pro Asn Leu Arg Asn Val Val Ala Leu 420 425 430 Asp Thr Glu Val Ala Ser Asn Arg Ile Tyr Trp Ser Asp Leu Ser Gln 435 440 445 Arg Met Ile Cys Ser Thr Gln Leu Asp Arg Ala His Gly Val Ser Ser 450 455 460 Tyr Asp Thr Val Ile Ser Arg Asp Ile Gln Ala Pro Asp Gly Leu Ala 465 470 475 480 Val Asp Trp Ile His Ser Asn Ile Tyr Trp Thr Asp Ser Val Leu Gly 485 490 495 Thr Val Ser Val Ala Asp Thr Lys Gly Val Lys Arg Lys Thr Leu Phe 500 505 510 Arg Glu Asn Gly Ser Lys Pro Arg Ala Ile Val Val Asp Pro Val His 515 520 525 Gly Phe Met Tyr Trp Thr Asp Trp Gly Thr Pro Ala Lys Ile Lys Lys 530 535 540 Gly Gly Leu Asn Gly Val Asp Ile Tyr Ser Leu Val Thr Glu Asn Ile 545 550 555 560 Gln Trp Pro Asn Gly Ile Thr Leu Asp Leu Leu Ser Gly Arg Leu Tyr 565 570 575 Trp Val Asp Ser Lys Leu His Ser Ile Ser Ser Ile Asp Val Asn Gly 580 585 590 Gly Asn Arg Lys Thr Ile Leu Glu Asp Glu Lys Arg Leu Ala His Pro 595 600 605 Phe Ser Leu Ala Val Phe Glu Asp Lys Val Phe Trp Thr Asp Ile Ile 610 615 620 Asn Glu Ala Ile Phe Ser Ala Asn Arg Leu Thr Gly Ser Asp Val Asn 625 630 635 640 Leu Leu Ala Glu Asn Leu Leu Ser Pro Glu Asp Met Val Leu Phe His 645 650 655 Asn Leu Thr Gln Pro Arg Gly Val Asn Trp Cys Glu Arg Thr Thr Leu 660 665 670 Ser Asn Gly Gly Cys Gln Tyr Leu Cys Leu Pro Ala Pro Gln Ile Asn 675 680 685 Pro His Ser Pro Lys Phe Thr Cys Ala Cys Pro Asp Gly Met Leu Leu 690 695 700 Ala Arg Asp Met Arg Ser Cys Leu Thr Glu Ala Glu Ala Ala Val Ala 705 710 715 720 Thr Gln Glu Thr Ser Thr Val Arg Leu Lys Val Ser Ser Thr Ala Val 725 730 735 Arg Thr Gln His Thr Thr Thr Arg Pro Val Pro Asp Thr Ser Arg Leu 740 745 750 Pro Gly Ala Thr Pro Gly Leu Thr Thr Val Glu Ile Val Thr Met Ser 755 760 765 His Gln Ala Leu Gly Asp Val Ala Gly Arg Gly Asn Glu Lys Lys Pro 770 775 780 Ser Ser Val Arg Ala Leu Ser Ile Val Leu Pro Ile Val Leu Leu Val 785 790 795 800 Phe Leu Cys Leu Gly Val Phe Leu Leu Trp Lys Asn Trp Arg Leu Lys 805 810 815 Asn Ile Asn Ser Ile Asn Phe Asp Asn Pro Val Tyr Gln Lys Thr Thr 820 825 830 Glu Asp Glu Val His Ile Cys His Asn Gln Asp Gly Tyr Ser Tyr Pro 835 840 845 Ser Arg Gln Met Val Ser Leu Glu Asp Asp Val Ala 850 855 860 68800PRTHomo sapiens 68Met Gly Thr Ser Ala Leu Trp Ala Leu Trp Leu Leu Leu Ala Leu Cys 1 5 10 15 Trp Ala Pro Arg Glu Ser Gly Ala Thr Gly Thr Gly Arg Lys Ala Lys 20 25 30 Cys Glu Pro Ser Gln Phe Gln Cys Thr Asn Gly Arg Cys Ile Thr Leu 35 40 45 Leu Trp Lys Cys Asp Gly Asp Glu Asp Cys Val Asp Gly Ser Asp Glu 50 55 60 Lys Asn Cys Val Lys Lys Thr Cys Ala Glu Ser Asp Phe Val Cys Asn 65 70 75 80 Asn Gly Gln Cys Val Pro Ser Arg Trp Lys Cys Asp Gly Asp Pro Asp 85 90 95 Cys Glu Asp Gly Ser Asp Glu Ser Pro Glu Gln Cys His Met Arg Thr 100 105 110 Cys Arg Ile His Glu Ile Ser Cys Gly Ala His Ser Thr Gln Cys Ile 115 120 125 Pro Val Ser Trp Arg Cys Asp Gly Glu Asn Asp Cys Asp Ser Gly Glu 130 135 140 Asp Glu Glu Asn Cys Gly Asn Ile Thr Cys Ser Pro Asp Glu Phe Thr 145 150 155 160 Cys Ser Ser Gly Arg Cys Ile Ser Arg Asn Phe Val Cys Asn Gly Gln 165 170 175 Asp Asp Cys Ser Asp Gly Ser Asp Glu Leu Asp Cys Ala Pro Pro Thr 180 185 190 Cys Gly Ala His Glu Phe Gln Cys Ser Thr Ser Ser Cys Ile Pro Ile 195 200 205 Ser Trp Val Cys Asp Asp Asp Ala Asp Cys Ser Asp Gln Ser Asp Glu 210 215 220 Ser Leu Glu Gln Cys Gly Arg Gln Pro Val Ile His Thr Lys Cys Pro 225 230 235 240 Ala Ser Glu Ile Gln Cys Gly Ser Gly Glu Cys Ile His Lys Lys Trp 245 250 255 Arg Cys Asp Gly Asp Pro Asp Cys Lys Asp Gly Ser Asp Glu Val Asn 260 265 270 Cys Pro Ser Arg Thr Cys Arg Pro Asp Gln Phe Glu Cys Glu Asp Gly 275 280 285 Ser Cys Ile His Gly Ser Arg Gln Cys Asn Gly Ile Arg Asp Cys Val 290 295 300 Asp Gly Ser Asp Glu Val Asn Cys Lys Asn Val Asn Gln Cys Leu Gly 305 310 315 320 Pro Gly Lys Phe Lys Cys Arg Ser Gly Glu Cys Ile Asp Ile Ser Lys 325 330 335 Val Cys Asn Gln Glu Gln Asp Cys Arg Asp Trp Ser Asp Glu Pro Leu 340 345 350 Lys Glu Cys His Ile Asn Glu Cys Leu Val Asn Asn Gly Gly Cys Ser 355 360 365 His Ile Cys Lys Asp Leu Val Ile Gly Tyr Glu Cys Asp Cys Ala Ala 370 375 380 Gly Phe Glu Leu Ile Asp Arg Lys Thr Cys Gly Asp Ile Asp Glu Cys 385 390 395 400 Gln Asn Pro Gly Ile Cys Ser Gln Ile Cys

Ile Asn Leu Lys Gly Gly 405 410 415 Tyr Lys Cys Glu Cys Ser Arg Gly Tyr Gln Met Asp Leu Ala Thr Gly 420 425 430 Val Cys Lys Ala Val Gly Lys Glu Pro Ser Leu Ile Phe Thr Asn Arg 435 440 445 Arg Asp Ile Arg Lys Ile Gly Leu Glu Arg Lys Glu Tyr Ile Gln Leu 450 455 460 Val Glu Gln Leu Arg Asn Thr Val Ala Leu Asp Ala Asp Ile Ala Ala 465 470 475 480 Gln Lys Leu Phe Trp Ala Asp Leu Ser Gln Lys Ala Ile Phe Ser Ala 485 490 495 Ser Ile Asp Asp Lys Val Gly Arg His Val Lys Met Ile Asp Asn Val 500 505 510 Tyr Asn Pro Ala Ala Ile Ala Val Asp Trp Val Tyr Lys Thr Ile Tyr 515 520 525 Trp Thr Asp Ala Ala Ser Lys Thr Ile Ser Val Ala Thr Leu Asp Gly 530 535 540 Thr Lys Arg Lys Phe Leu Phe Asn Ser Asp Leu Arg Glu Pro Ala Ser 545 550 555 560 Ile Ala Val Asp Pro Leu Ser Gly Phe Val Tyr Trp Ser Asp Trp Gly 565 570 575 Glu Pro Ala Lys Ile Glu Lys Ala Gly Met Asn Gly Phe Asp Arg Arg 580 585 590 Pro Leu Val Thr Ala Asp Ile Gln Trp Pro Asn Gly Ile Thr Leu Asp 595 600 605 Leu Ile Lys Ser Arg Leu Tyr Trp Leu Asp Ser Lys Leu His Met Leu 610 615 620 Ser Ser Val Asp Leu Asn Gly Gln Asp Arg Arg Ile Val Leu Lys Ser 625 630 635 640 Leu Glu Phe Leu Ala His Pro Leu Ala Leu Thr Ile Phe Glu Asp Arg 645 650 655 Val Tyr Trp Ile Asp Gly Glu Asn Glu Ala Val Tyr Gly Ala Asn Lys 660 665 670 Phe Thr Gly Ser Glu Leu Ala Thr Leu Val Asn Asn Leu Asn Asp Ala 675 680 685 Gln Asp Ile Ile Val Tyr His Glu Leu Val Gln Pro Ser Gly Lys Asn 690 695 700 Trp Cys Glu Glu Asp Met Glu Asn Gly Gly Cys Glu Tyr Leu Cys Leu 705 710 715 720 Pro Ala Pro Gln Ile Asn Asp His Ser Pro Lys Tyr Thr Cys Ser Cys 725 730 735 Pro Ser Gly Tyr Asn Val Glu Glu Asn Gly Arg Asp Cys Gln Arg Ile 740 745 750 Asn Val Thr Thr Ala Val Ser Glu Val Ser Val Pro Pro Lys Gly Thr 755 760 765 Ser Ala Ala Trp Ala Ile Leu Pro Leu Leu Leu Leu Val Met Ala Ala 770 775 780 Val Gly Gly Tyr Leu Met Trp Arg Asn Trp Gln His Lys Asn Met Lys 785 790 795 800 69873PRTHomo sapiens 69Met Gly Thr Ser Ala Leu Trp Ala Leu Trp Leu Leu Leu Ala Leu Cys 1 5 10 15 Trp Ala Pro Arg Glu Ser Gly Ala Thr Gly Thr Gly Arg Lys Ala Lys 20 25 30 Cys Glu Pro Ser Gln Phe Gln Cys Thr Asn Gly Arg Cys Ile Thr Leu 35 40 45 Leu Trp Lys Cys Asp Gly Asp Glu Asp Cys Val Asp Gly Ser Asp Glu 50 55 60 Lys Asn Cys Val Lys Lys Thr Cys Ala Glu Ser Asp Phe Val Cys Asn 65 70 75 80 Asn Gly Gln Cys Val Pro Ser Arg Trp Lys Cys Asp Gly Asp Pro Asp 85 90 95 Cys Glu Asp Gly Ser Asp Glu Ser Pro Glu Gln Cys His Met Arg Thr 100 105 110 Cys Arg Ile His Glu Ile Ser Cys Gly Ala His Ser Thr Gln Cys Ile 115 120 125 Pro Val Ser Trp Arg Cys Asp Gly Glu Asn Asp Cys Asp Ser Gly Glu 130 135 140 Asp Glu Glu Asn Cys Gly Asn Ile Thr Cys Ser Pro Asp Glu Phe Thr 145 150 155 160 Cys Ser Ser Gly Arg Cys Ile Ser Arg Asn Phe Val Cys Asn Gly Gln 165 170 175 Asp Asp Cys Ser Asp Gly Ser Asp Glu Leu Asp Cys Ala Pro Pro Thr 180 185 190 Cys Gly Ala His Glu Phe Gln Cys Ser Thr Ser Ser Cys Ile Pro Ile 195 200 205 Ser Trp Val Cys Asp Asp Asp Ala Asp Cys Ser Asp Gln Ser Asp Glu 210 215 220 Ser Leu Glu Gln Cys Gly Arg Gln Pro Val Ile His Thr Lys Cys Pro 225 230 235 240 Ala Ser Glu Ile Gln Cys Gly Ser Gly Glu Cys Ile His Lys Lys Trp 245 250 255 Arg Cys Asp Gly Asp Pro Asp Cys Lys Asp Gly Ser Asp Glu Val Asn 260 265 270 Cys Pro Ser Arg Thr Cys Arg Pro Asp Gln Phe Glu Cys Glu Asp Gly 275 280 285 Ser Cys Ile His Gly Ser Arg Gln Cys Asn Gly Ile Arg Asp Cys Val 290 295 300 Asp Gly Ser Asp Glu Val Asn Cys Lys Asn Val Asn Gln Cys Leu Gly 305 310 315 320 Pro Gly Lys Phe Lys Cys Arg Ser Gly Glu Cys Ile Asp Ile Ser Lys 325 330 335 Val Cys Asn Gln Glu Gln Asp Cys Arg Asp Trp Ser Asp Glu Pro Leu 340 345 350 Lys Glu Cys His Ile Asn Glu Cys Leu Val Asn Asn Gly Gly Cys Ser 355 360 365 His Ile Cys Lys Asp Leu Val Ile Gly Tyr Glu Cys Asp Cys Ala Ala 370 375 380 Gly Phe Glu Leu Ile Asp Arg Lys Thr Cys Gly Asp Ile Asp Glu Cys 385 390 395 400 Gln Asn Pro Gly Ile Cys Ser Gln Ile Cys Ile Asn Leu Lys Gly Gly 405 410 415 Tyr Lys Cys Glu Cys Ser Arg Gly Tyr Gln Met Asp Leu Ala Thr Gly 420 425 430 Val Cys Lys Ala Val Gly Lys Glu Pro Ser Leu Ile Phe Thr Asn Arg 435 440 445 Arg Asp Ile Arg Lys Ile Gly Leu Glu Arg Lys Glu Tyr Ile Gln Leu 450 455 460 Val Glu Gln Leu Arg Asn Thr Val Ala Leu Asp Ala Asp Ile Ala Ala 465 470 475 480 Gln Lys Leu Phe Trp Ala Asp Leu Ser Gln Lys Ala Ile Phe Ser Ala 485 490 495 Ser Ile Asp Asp Lys Val Gly Arg His Val Lys Met Ile Asp Asn Val 500 505 510 Tyr Asn Pro Ala Ala Ile Ala Val Asp Trp Val Tyr Lys Thr Ile Tyr 515 520 525 Trp Thr Asp Ala Ala Ser Lys Thr Ile Ser Val Ala Thr Leu Asp Gly 530 535 540 Thr Lys Arg Lys Phe Leu Phe Asn Ser Asp Leu Arg Glu Pro Ala Ser 545 550 555 560 Ile Ala Val Asp Pro Leu Ser Gly Phe Val Tyr Trp Ser Asp Trp Gly 565 570 575 Glu Pro Ala Lys Ile Glu Lys Ala Gly Met Asn Gly Phe Asp Arg Arg 580 585 590 Pro Leu Val Thr Ala Asp Ile Gln Trp Pro Asn Gly Ile Thr Leu Asp 595 600 605 Leu Ile Lys Ser Arg Leu Tyr Trp Leu Asp Ser Lys Leu His Met Leu 610 615 620 Ser Ser Val Asp Leu Asn Gly Gln Asp Arg Arg Ile Val Leu Lys Ser 625 630 635 640 Leu Glu Phe Leu Ala His Pro Leu Ala Leu Thr Ile Phe Glu Asp Arg 645 650 655 Val Tyr Trp Ile Asp Gly Glu Asn Glu Ala Val Tyr Gly Ala Asn Lys 660 665 670 Phe Thr Gly Ser Glu Leu Ala Thr Leu Val Asn Asn Leu Asn Asp Ala 675 680 685 Gln Asp Ile Ile Val Tyr His Glu Leu Val Gln Pro Ser Gly Lys Asn 690 695 700 Trp Cys Glu Glu Asp Met Glu Asn Gly Gly Cys Glu Tyr Leu Cys Leu 705 710 715 720 Pro Ala Pro Gln Ile Asn Asp His Ser Pro Lys Tyr Thr Cys Ser Cys 725 730 735 Pro Ser Gly Tyr Asn Val Glu Glu Asn Gly Arg Asp Cys Gln Ser Thr 740 745 750 Ala Thr Thr Val Thr Tyr Ser Glu Thr Lys Asp Thr Asn Thr Thr Glu 755 760 765 Ile Ser Ala Thr Ser Gly Leu Val Pro Gly Gly Ile Asn Val Thr Thr 770 775 780 Ala Val Ser Glu Val Ser Val Pro Pro Lys Gly Thr Ser Ala Ala Trp 785 790 795 800 Ala Ile Leu Pro Leu Leu Leu Leu Val Met Ala Ala Val Gly Gly Tyr 805 810 815 Leu Met Trp Arg Asn Trp Gln His Lys Asn Met Lys Ser Met Asn Phe 820 825 830 Asp Asn Pro Val Tyr Leu Lys Thr Thr Glu Glu Asp Leu Ser Ile Asp 835 840 845 Ile Gly Arg His Ser Ala Ser Val Gly His Thr Tyr Pro Ala Ile Ser 850 855 860 Val Val Ser Thr Asp Asp Asp Leu Ala 865 870 701255PRTHomo sapiens 70Met Glu Leu Ala Ala Leu Cys Arg Trp Gly Leu Leu Leu Ala Leu Leu 1 5 10 15 Pro Pro Gly Ala Ala Ser Thr Gln Val Cys Thr Gly Thr Asp Met Lys 20 25 30 Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu Arg His 35 40 45 Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu Thr Tyr 50 55 60 Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln Glu Val 65 70 75 80 Gln Gly Tyr Val Leu Ile Ala His Asn Gln Val Arg Gln Val Pro Leu 85 90 95 Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp Asn Tyr 100 105 110 Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr Thr Pro 115 120 125 Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu Arg Ser 130 135 140 Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn Pro Gln 145 150 155 160 Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His Lys Asn 165 170 175 Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg Ala Cys 180 185 190 His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly Glu Ser 195 200 205 Ser Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly Gly Cys 210 215 220 Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu Gln Cys 225 230 235 240 Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala Cys Leu 245 250 255 His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala Leu Val 260 265 270 Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu Gly Arg 275 280 285 Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn Tyr Leu 290 295 300 Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu His Asn Gln 305 310 315 320 Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys Ser Lys 325 330 335 Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu Arg Glu 340 345 350 Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly Cys Lys 355 360 365 Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp Gly Asp 370 375 380 Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln Val Phe 385 390 395 400 Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala Trp Pro 405 410 415 Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln Val Ile Arg 420 425 430 Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu Thr Leu Gln Gly Leu 435 440 445 Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly Ser Gly 450 455 460 Leu Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His Thr Val 465 470 475 480 Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu His Thr 485 490 495 Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala Cys His 500 505 510 Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr Gln Cys 515 520 525 Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu Glu Cys 530 535 540 Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg His Cys 545 550 555 560 Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val Thr Cys 565 570 575 Phe Gly Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr Lys Asp 580 585 590 Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro Asp Leu 595 600 605 Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala Cys Gln 610 615 620 Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp Asp Lys 625 630 635 640 Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu Thr Ser Ile Ile Ser 645 650 655 Ala Val Val Gly Ile Leu Leu Val Val Val Leu Gly Val Val Phe Gly 660 665 670 Ile Leu Ile Lys Arg Arg Gln Gln Lys Ile Arg Lys Tyr Thr Met Arg 675 680 685 Arg Leu Leu Gln Glu Thr Glu Leu Val Glu Pro Leu Thr Pro Ser Gly 690 695 700 Ala Met Pro Asn Gln Ala Gln Met Arg Ile Leu Lys Glu Thr Glu Leu 705 710 715 720 Arg Lys Val Lys Val Leu Gly Ser Gly Ala Phe Gly Thr Val Tyr Lys 725 730 735 Gly Ile Trp Ile Pro Asp Gly Glu Asn Val Lys Ile Pro Val Ala Ile 740 745 750 Lys Val Leu Arg Glu Asn Thr Ser Pro Lys Ala Asn Lys Glu Ile Leu 755 760 765 Asp Glu Ala Tyr Val Met Ala Gly Val Gly Ser Pro Tyr Val Ser Arg 770 775 780 Leu Leu Gly Ile Cys Leu Thr Ser Thr Val Gln Leu Val Thr Gln Leu 785 790 795 800 Met Pro Tyr Gly Cys Leu Leu Asp His Val Arg Glu Asn Arg Gly Arg 805 810 815 Leu Gly Ser Gln Asp Leu Leu Asn Trp Cys Met Gln Ile Ala Lys Gly 820 825 830 Met Ser Tyr Leu Glu Asp Val Arg Leu Val His Arg Asp Leu Ala Ala 835 840 845 Arg Asn Val Leu Val Lys Ser Pro Asn His Val Lys Ile Thr Asp Phe 850 855 860 Gly Leu Ala Arg Leu Leu Asp Ile Asp Glu Thr Glu Tyr His Ala Asp 865 870 875 880 Gly Gly Lys Val Pro Ile Lys Trp Met Ala Leu Glu Ser Ile Leu Arg 885 890 895 Arg Arg Phe Thr His Gln Ser Asp Val Trp Ser Tyr Gly Val Thr Val 900 905 910 Trp Glu Leu Met Thr Phe Gly Ala Lys Pro Tyr Asp Gly Ile Pro Ala 915 920 925 Arg Glu Ile Pro Asp Leu Leu Glu Lys Gly Glu Arg Leu Pro Gln Pro 930 935 940 Pro Ile Cys Thr Ile Asp Val Tyr Met Ile Met Val Lys Cys Trp Met 945 950 955 960 Ile Asp Ser Glu Cys Arg Pro Arg Phe Arg Glu Leu Val Ser Glu Phe 965 970 975 Ser Arg Met Ala Arg Asp Pro Gln Arg Phe Val Val Ile Gln Asn Glu 980 985 990 Asp Leu Gly Pro Ala Ser Pro Leu Asp Ser Thr Phe Tyr Arg Ser Leu 995 1000 1005 Leu Glu Asp Asp Asp Met Gly Asp Leu Val Asp Ala Glu Glu Tyr 1010 1015 1020 Leu Val Pro Gln Gln Gly Phe Phe Cys Pro Asp Pro Ala Pro Gly

1025 1030 1035 Ala Gly Gly Met Val His His Arg His Arg Ser Ser Ser Thr Arg 1040 1045 1050 Ser Gly Gly Gly Asp Leu Thr Leu Gly Leu Glu Pro Ser Glu Glu 1055 1060 1065 Glu Ala Pro Arg Ser Pro Leu Ala Pro Ser Glu Gly Ala Gly Ser 1070 1075 1080 Asp Val Phe Asp Gly Asp Leu Gly Met Gly Ala Ala Lys Gly Leu 1085 1090 1095 Gln Ser Leu Pro Thr His Asp Pro Ser Pro Leu Gln Arg Tyr Ser 1100 1105 1110 Glu Asp Pro Thr Val Pro Leu Pro Ser Glu Thr Asp Gly Tyr Val 1115 1120 1125 Ala Pro Leu Thr Cys Ser Pro Gln Pro Glu Tyr Val Asn Gln Pro 1130 1135 1140 Asp Val Arg Pro Gln Pro Pro Ser Pro Arg Glu Gly Pro Leu Pro 1145 1150 1155 Ala Ala Arg Pro Ala Gly Ala Thr Leu Glu Arg Pro Lys Thr Leu 1160 1165 1170 Ser Pro Gly Lys Asn Gly Val Val Lys Asp Val Phe Ala Phe Gly 1175 1180 1185 Gly Ala Val Glu Asn Pro Glu Tyr Leu Thr Pro Gln Gly Gly Ala 1190 1195 1200 Ala Pro Gln Pro His Pro Pro Pro Ala Phe Ser Pro Ala Phe Asp 1205 1210 1215 Asn Leu Tyr Tyr Trp Asp Gln Asp Pro Pro Glu Arg Gly Ala Pro 1220 1225 1230 Pro Ser Thr Phe Lys Gly Thr Pro Thr Ala Glu Asn Pro Glu Tyr 1235 1240 1245 Leu Gly Leu Asp Val Pro Val 1250 1255 711225PRTHomo sapiens 71Met Lys Leu Arg Leu Pro Ala Ser Pro Glu Thr His Leu Asp Met Leu 1 5 10 15 Arg His Leu Tyr Gln Gly Cys Gln Val Val Gln Gly Asn Leu Glu Leu 20 25 30 Thr Tyr Leu Pro Thr Asn Ala Ser Leu Ser Phe Leu Gln Asp Ile Gln 35 40 45 Glu Val Gln Gly Tyr Val Leu Ile Ala His Asn Gln Val Arg Gln Val 50 55 60 Pro Leu Gln Arg Leu Arg Ile Val Arg Gly Thr Gln Leu Phe Glu Asp 65 70 75 80 Asn Tyr Ala Leu Ala Val Leu Asp Asn Gly Asp Pro Leu Asn Asn Thr 85 90 95 Thr Pro Val Thr Gly Ala Ser Pro Gly Gly Leu Arg Glu Leu Gln Leu 100 105 110 Arg Ser Leu Thr Glu Ile Leu Lys Gly Gly Val Leu Ile Gln Arg Asn 115 120 125 Pro Gln Leu Cys Tyr Gln Asp Thr Ile Leu Trp Lys Asp Ile Phe His 130 135 140 Lys Asn Asn Gln Leu Ala Leu Thr Leu Ile Asp Thr Asn Arg Ser Arg 145 150 155 160 Ala Cys His Pro Cys Ser Pro Met Cys Lys Gly Ser Arg Cys Trp Gly 165 170 175 Glu Ser Ser Glu Asp Cys Gln Ser Leu Thr Arg Thr Val Cys Ala Gly 180 185 190 Gly Cys Ala Arg Cys Lys Gly Pro Leu Pro Thr Asp Cys Cys His Glu 195 200 205 Gln Cys Ala Ala Gly Cys Thr Gly Pro Lys His Ser Asp Cys Leu Ala 210 215 220 Cys Leu His Phe Asn His Ser Gly Ile Cys Glu Leu His Cys Pro Ala 225 230 235 240 Leu Val Thr Tyr Asn Thr Asp Thr Phe Glu Ser Met Pro Asn Pro Glu 245 250 255 Gly Arg Tyr Thr Phe Gly Ala Ser Cys Val Thr Ala Cys Pro Tyr Asn 260 265 270 Tyr Leu Ser Thr Asp Val Gly Ser Cys Thr Leu Val Cys Pro Leu His 275 280 285 Asn Gln Glu Val Thr Ala Glu Asp Gly Thr Gln Arg Cys Glu Lys Cys 290 295 300 Ser Lys Pro Cys Ala Arg Val Cys Tyr Gly Leu Gly Met Glu His Leu 305 310 315 320 Arg Glu Val Arg Ala Val Thr Ser Ala Asn Ile Gln Glu Phe Ala Gly 325 330 335 Cys Lys Lys Ile Phe Gly Ser Leu Ala Phe Leu Pro Glu Ser Phe Asp 340 345 350 Gly Asp Pro Ala Ser Asn Thr Ala Pro Leu Gln Pro Glu Gln Leu Gln 355 360 365 Val Phe Glu Thr Leu Glu Glu Ile Thr Gly Tyr Leu Tyr Ile Ser Ala 370 375 380 Trp Pro Asp Ser Leu Pro Asp Leu Ser Val Phe Gln Asn Leu Gln Val 385 390 395 400 Ile Arg Gly Arg Ile Leu His Asn Gly Ala Tyr Ser Leu Thr Leu Gln 405 410 415 Gly Leu Gly Ile Ser Trp Leu Gly Leu Arg Ser Leu Arg Glu Leu Gly 420 425 430 Ser Gly Leu Ala Leu Ile His His Asn Thr His Leu Cys Phe Val His 435 440 445 Thr Val Pro Trp Asp Gln Leu Phe Arg Asn Pro His Gln Ala Leu Leu 450 455 460 His Thr Ala Asn Arg Pro Glu Asp Glu Cys Val Gly Glu Gly Leu Ala 465 470 475 480 Cys His Gln Leu Cys Ala Arg Gly His Cys Trp Gly Pro Gly Pro Thr 485 490 495 Gln Cys Val Asn Cys Ser Gln Phe Leu Arg Gly Gln Glu Cys Val Glu 500 505 510 Glu Cys Arg Val Leu Gln Gly Leu Pro Arg Glu Tyr Val Asn Ala Arg 515 520 525 His Cys Leu Pro Cys His Pro Glu Cys Gln Pro Gln Asn Gly Ser Val 530 535 540 Thr Cys Phe Gly Pro Glu Ala Asp Gln Cys Val Ala Cys Ala His Tyr 545 550 555 560 Lys Asp Pro Pro Phe Cys Val Ala Arg Cys Pro Ser Gly Val Lys Pro 565 570 575 Asp Leu Ser Tyr Met Pro Ile Trp Lys Phe Pro Asp Glu Glu Gly Ala 580 585 590 Cys Gln Pro Cys Pro Ile Asn Cys Thr His Ser Cys Val Asp Leu Asp 595 600 605 Asp Lys Gly Cys Pro Ala Glu Gln Arg Ala Ser Pro Leu Thr Ser Ile 610 615 620 Ile Ser Ala Val Val Gly Ile Leu Leu Val Val Val Leu Gly Val Val 625 630 635 640 Phe Gly Ile Leu Ile Lys Arg Arg Gln Gln Lys Ile Arg Lys Tyr Thr 645 650 655 Met Arg Arg Leu Leu Gln Glu Thr Glu Leu Val Glu Pro Leu Thr Pro 660 665 670 Ser Gly Ala Met Pro Asn Gln Ala Gln Met Arg Ile Leu Lys Glu Thr 675 680 685 Glu Leu Arg Lys Val Lys Val Leu Gly Ser Gly Ala Phe Gly Thr Val 690 695 700 Tyr Lys Gly Ile Trp Ile Pro Asp Gly Glu Asn Val Lys Ile Pro Val 705 710 715 720 Ala Ile Lys Val Leu Arg Glu Asn Thr Ser Pro Lys Ala Asn Lys Glu 725 730 735 Ile Leu Asp Glu Ala Tyr Val Met Ala Gly Val Gly Ser Pro Tyr Val 740 745 750 Ser Arg Leu Leu Gly Ile Cys Leu Thr Ser Thr Val Gln Leu Val Thr 755 760 765 Gln Leu Met Pro Tyr Gly Cys Leu Leu Asp His Val Arg Glu Asn Arg 770 775 780 Gly Arg Leu Gly Ser Gln Asp Leu Leu Asn Trp Cys Met Gln Ile Ala 785 790 795 800 Lys Gly Met Ser Tyr Leu Glu Asp Val Arg Leu Val His Arg Asp Leu 805 810 815 Ala Ala Arg Asn Val Leu Val Lys Ser Pro Asn His Val Lys Ile Thr 820 825 830 Asp Phe Gly Leu Ala Arg Leu Leu Asp Ile Asp Glu Thr Glu Tyr His 835 840 845 Ala Asp Gly Gly Lys Val Pro Ile Lys Trp Met Ala Leu Glu Ser Ile 850 855 860 Leu Arg Arg Arg Phe Thr His Gln Ser Asp Val Trp Ser Tyr Gly Val 865 870 875 880 Thr Val Trp Glu Leu Met Thr Phe Gly Ala Lys Pro Tyr Asp Gly Ile 885 890 895 Pro Ala Arg Glu Ile Pro Asp Leu Leu Glu Lys Gly Glu Arg Leu Pro 900 905 910 Gln Pro Pro Ile Cys Thr Ile Asp Val Tyr Met Ile Met Val Lys Cys 915 920 925 Trp Met Ile Asp Ser Glu Cys Arg Pro Arg Phe Arg Glu Leu Val Ser 930 935 940 Glu Phe Ser Arg Met Ala Arg Asp Pro Gln Arg Phe Val Val Ile Gln 945 950 955 960 Asn Glu Asp Leu Gly Pro Ala Ser Pro Leu Asp Ser Thr Phe Tyr Arg 965 970 975 Ser Leu Leu Glu Asp Asp Asp Met Gly Asp Leu Val Asp Ala Glu Glu 980 985 990 Tyr Leu Val Pro Gln Gln Gly Phe Phe Cys Pro Asp Pro Ala Pro Gly 995 1000 1005 Ala Gly Gly Met Val His His Arg His Arg Ser Ser Ser Thr Arg 1010 1015 1020 Ser Gly Gly Gly Asp Leu Thr Leu Gly Leu Glu Pro Ser Glu Glu 1025 1030 1035 Glu Ala Pro Arg Ser Pro Leu Ala Pro Ser Glu Gly Ala Gly Ser 1040 1045 1050 Asp Val Phe Asp Gly Asp Leu Gly Met Gly Ala Ala Lys Gly Leu 1055 1060 1065 Gln Ser Leu Pro Thr His Asp Pro Ser Pro Leu Gln Arg Tyr Ser 1070 1075 1080 Glu Asp Pro Thr Val Pro Leu Pro Ser Glu Thr Asp Gly Tyr Val 1085 1090 1095 Ala Pro Leu Thr Cys Ser Pro Gln Pro Glu Tyr Val Asn Gln Pro 1100 1105 1110 Asp Val Arg Pro Gln Pro Pro Ser Pro Arg Glu Gly Pro Leu Pro 1115 1120 1125 Ala Ala Arg Pro Ala Gly Ala Thr Leu Glu Arg Pro Lys Thr Leu 1130 1135 1140 Ser Pro Gly Lys Asn Gly Val Val Lys Asp Val Phe Ala Phe Gly 1145 1150 1155 Gly Ala Val Glu Asn Pro Glu Tyr Leu Thr Pro Gln Gly Gly Ala 1160 1165 1170 Ala Pro Gln Pro His Pro Pro Pro Ala Phe Ser Pro Ala Phe Asp 1175 1180 1185 Asn Leu Tyr Tyr Trp Asp Gln Asp Pro Pro Glu Arg Gly Ala Pro 1190 1195 1200 Pro Ser Thr Phe Lys Gly Thr Pro Thr Ala Glu Asn Pro Glu Tyr 1205 1210 1215 Leu Gly Leu Asp Val Pro Val 1220 1225 72654PRTHomo sapiens 72Met Lys Leu Ser Leu Val Ala Ala Met Leu Leu Leu Leu Ser Ala Ala 1 5 10 15 Arg Ala Glu Glu Glu Asp Lys Lys Glu Asp Val Gly Thr Val Val Gly 20 25 30 Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val Gly Val Phe Lys Asn Gly 35 40 45 Arg Val Glu Ile Ile Ala Asn Asp Gln Gly Asn Arg Ile Thr Pro Ser 50 55 60 Tyr Val Ala Phe Thr Pro Glu Gly Glu Arg Leu Ile Gly Asp Ala Ala 65 70 75 80 Lys Asn Gln Leu Thr Ser Asn Pro Glu Asn Thr Val Phe Asp Ala Lys 85 90 95 Arg Leu Ile Gly Arg Thr Trp Asn Asp Pro Ser Val Gln Gln Asp Ile 100 105 110 Lys Phe Leu Pro Phe Lys Val Val Glu Lys Lys Thr Lys Pro Tyr Ile 115 120 125 Gln Val Asp Ile Gly Gly Gly Gln Thr Lys Thr Phe Ala Pro Glu Glu 130 135 140 Ile Ser Ala Met Val Leu Thr Lys Met Lys Glu Thr Ala Glu Ala Tyr 145 150 155 160 Leu Gly Lys Lys Val Thr His Ala Val Val Thr Val Pro Ala Tyr Phe 165 170 175 Asn Asp Ala Gln Arg Gln Ala Thr Lys Asp Ala Gly Thr Ile Ala Gly 180 185 190 Leu Asn Val Met Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile Ala 195 200 205 Tyr Gly Leu Asp Lys Arg Glu Gly Glu Lys Asn Ile Leu Val Phe Asp 210 215 220 Leu Gly Gly Gly Thr Phe Asp Val Ser Leu Leu Thr Ile Asp Asn Gly 225 230 235 240 Val Phe Glu Val Val Ala Thr Asn Gly Asp Thr His Leu Gly Gly Glu 245 250 255 Asp Phe Asp Gln Arg Val Met Glu His Phe Ile Lys Leu Tyr Lys Lys 260 265 270 Lys Thr Gly Lys Asp Val Arg Lys Asp Asn Arg Ala Val Gln Lys Leu 275 280 285 Arg Arg Glu Val Glu Lys Ala Lys Arg Ala Leu Ser Ser Gln His Gln 290 295 300 Ala Arg Ile Glu Ile Glu Ser Phe Tyr Glu Gly Glu Asp Phe Ser Glu 305 310 315 320 Thr Leu Thr Arg Ala Lys Phe Glu Glu Leu Asn Met Asp Leu Phe Arg 325 330 335 Ser Thr Met Lys Pro Val Gln Lys Val Leu Glu Asp Ser Asp Leu Lys 340 345 350 Lys Ser Asp Ile Asp Glu Ile Val Leu Val Gly Gly Ser Thr Arg Ile 355 360 365 Pro Lys Ile Gln Gln Leu Val Lys Glu Phe Phe Asn Gly Lys Glu Pro 370 375 380 Ser Arg Gly Ile Asn Pro Asp Glu Ala Val Ala Tyr Gly Ala Ala Val 385 390 395 400 Gln Ala Gly Val Leu Ser Gly Asp Gln Asp Thr Gly Asp Leu Val Leu 405 410 415 Leu Asp Val Cys Pro Leu Thr Leu Gly Ile Glu Thr Val Gly Gly Val 420 425 430 Met Thr Lys Leu Ile Pro Arg Asn Thr Val Val Pro Thr Lys Lys Ser 435 440 445 Gln Ile Phe Ser Thr Ala Ser Asp Asn Gln Pro Thr Val Thr Ile Lys 450 455 460 Val Tyr Glu Gly Glu Arg Pro Leu Thr Lys Asp Asn His Leu Leu Gly 465 470 475 480 Thr Phe Asp Leu Thr Gly Ile Pro Pro Ala Pro Arg Gly Val Pro Gln 485 490 495 Ile Glu Val Thr Phe Glu Ile Asp Val Asn Gly Ile Leu Arg Val Thr 500 505 510 Ala Glu Asp Lys Gly Thr Gly Asn Lys Asn Lys Ile Thr Ile Thr Asn 515 520 525 Asp Gln Asn Arg Leu Thr Pro Glu Glu Ile Glu Arg Met Val Asn Asp 530 535 540 Ala Glu Lys Phe Ala Glu Glu Asp Lys Lys Leu Lys Glu Arg Ile Asp 545 550 555 560 Thr Arg Asn Glu Leu Glu Ser Tyr Ala Tyr Ser Leu Lys Asn Gln Ile 565 570 575 Gly Asp Lys Glu Lys Leu Gly Gly Lys Leu Ser Ser Glu Asp Lys Glu 580 585 590 Thr Met Glu Lys Ala Val Glu Glu Lys Ile Glu Trp Leu Glu Ser His 595 600 605 Gln Asp Ala Asp Ile Glu Asp Phe Lys Ala Lys Lys Lys Glu Leu Glu 610 615 620 Glu Ile Val Gln Pro Ile Ile Ser Lys Leu Tyr Gly Ser Ala Gly Pro 625 630 635 640 Pro Pro Thr Gly Glu Glu Asp Thr Ala Glu Lys Asp Glu Leu 645 650 73493PRTHomo sapiens 73Met Ser Lys Gly Pro Ala Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser 1 5 10 15 Cys Val Gly Val Phe Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp 20 25 30 Gln Gly Asn Arg Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu 35 40 45 Arg Leu Ile Gly Asp Ala Ala Lys Asn Gln Val Ala Met Asn Pro Thr 50 55 60 Asn Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Arg Phe Asp Asp 65 70 75 80 Ala Val Val Gln Ser Asp Met Lys His Trp Pro Phe Met Val Val Asn 85 90 95 Asp Ala Gly Arg Pro Lys Val Gln Val Glu Tyr Lys Gly Glu Thr Lys 100 105 110 Ser Phe Tyr Pro Glu Glu Val Ser Ser Met Val Leu Thr Lys Met Lys 115 120 125 Glu Ile Ala Glu Ala Tyr Leu Gly Lys Thr Val Thr Asn Ala Val Val 130 135 140 Thr Val Pro Ala Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp 145 150 155 160 Ala Gly Thr Ile Ala Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro 165

170 175 Thr Ala Ala Ala Ile Ala Tyr Gly Leu Asp Lys Lys Val Gly Ala Glu 180 185 190 Arg Asn Val Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser 195 200 205 Ile Leu Thr Ile Glu Asp Gly Ile Phe Glu Val Lys Ser Thr Ala Gly 210 215 220 Asp Thr His Leu Gly Gly Glu Asp Phe Asp Asn Arg Met Val Asn His 225 230 235 240 Phe Ile Ala Glu Phe Lys Arg Lys His Lys Lys Asp Ile Ser Glu Asn 245 250 255 Lys Arg Ala Val Arg Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg 260 265 270 Thr Leu Ser Ser Ser Thr Gln Ala Ser Ile Glu Ile Asp Ser Leu Tyr 275 280 285 Glu Gly Ile Asp Phe Tyr Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu 290 295 300 Leu Asn Ala Asp Leu Phe Arg Gly Thr Leu Asp Pro Val Glu Lys Ala 305 310 315 320 Leu Arg Asp Ala Lys Leu Asp Lys Ser Gln Ile His Asp Ile Val Leu 325 330 335 Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln Lys Leu Leu Gln Asp 340 345 350 Phe Phe Asn Gly Lys Glu Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala 355 360 365 Val Ala Tyr Gly Ala Ala Val Gln Ala Ala Ile Leu Ser Gly Asp Lys 370 375 380 Ser Glu Asn Val Gln Asp Leu Leu Leu Leu Asp Val Thr Pro Leu Ser 385 390 395 400 Leu Gly Ile Glu Thr Ala Gly Gly Val Met Thr Val Leu Ile Lys Arg 405 410 415 Asn Thr Thr Ile Pro Thr Lys Gln Thr Gln Thr Phe Thr Thr Tyr Ser 420 425 430 Asp Asn Gln Pro Gly Val Leu Ile Gln Val Tyr Glu Gly Glu Arg Ala 435 440 445 Met Thr Lys Asp Asn Asn Leu Leu Gly Lys Phe Glu Leu Thr Gly Met 450 455 460 Pro Gly Gly Met Pro Gly Gly Phe Pro Gly Gly Gly Ala Pro Pro Ser 465 470 475 480 Gly Gly Ala Ser Ser Gly Pro Thr Ile Glu Glu Val Asp 485 490 74646PRTHomo sapiens 74Met Ser Lys Gly Pro Ala Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser 1 5 10 15 Cys Val Gly Val Phe Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp 20 25 30 Gln Gly Asn Arg Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu 35 40 45 Arg Leu Ile Gly Asp Ala Ala Lys Asn Gln Val Ala Met Asn Pro Thr 50 55 60 Asn Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Arg Phe Asp Asp 65 70 75 80 Ala Val Val Gln Ser Asp Met Lys His Trp Pro Phe Met Val Val Asn 85 90 95 Asp Ala Gly Arg Pro Lys Val Gln Val Glu Tyr Lys Gly Glu Thr Lys 100 105 110 Ser Phe Tyr Pro Glu Glu Val Ser Ser Met Val Leu Thr Lys Met Lys 115 120 125 Glu Ile Ala Glu Ala Tyr Leu Gly Lys Thr Val Thr Asn Ala Val Val 130 135 140 Thr Val Pro Ala Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp 145 150 155 160 Ala Gly Thr Ile Ala Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro 165 170 175 Thr Ala Ala Ala Ile Ala Tyr Gly Leu Asp Lys Lys Val Gly Ala Glu 180 185 190 Arg Asn Val Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser 195 200 205 Ile Leu Thr Ile Glu Asp Gly Ile Phe Glu Val Lys Ser Thr Ala Gly 210 215 220 Asp Thr His Leu Gly Gly Glu Asp Phe Asp Asn Arg Met Val Asn His 225 230 235 240 Phe Ile Ala Glu Phe Lys Arg Lys His Lys Lys Asp Ile Ser Glu Asn 245 250 255 Lys Arg Ala Val Arg Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg 260 265 270 Thr Leu Ser Ser Ser Thr Gln Ala Ser Ile Glu Ile Asp Ser Leu Tyr 275 280 285 Glu Gly Ile Asp Phe Tyr Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu 290 295 300 Leu Asn Ala Asp Leu Phe Arg Gly Thr Leu Asp Pro Val Glu Lys Ala 305 310 315 320 Leu Arg Asp Ala Lys Leu Asp Lys Ser Gln Ile His Asp Ile Val Leu 325 330 335 Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln Lys Leu Leu Gln Asp 340 345 350 Phe Phe Asn Gly Lys Glu Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala 355 360 365 Val Ala Tyr Gly Ala Ala Val Gln Ala Ala Ile Leu Ser Gly Asp Lys 370 375 380 Ser Glu Asn Val Gln Asp Leu Leu Leu Leu Asp Val Thr Pro Leu Ser 385 390 395 400 Leu Gly Ile Glu Thr Ala Gly Gly Val Met Thr Val Leu Ile Lys Arg 405 410 415 Asn Thr Thr Ile Pro Thr Lys Gln Thr Gln Thr Phe Thr Thr Tyr Ser 420 425 430 Asp Asn Gln Pro Gly Val Leu Ile Gln Val Tyr Glu Gly Glu Arg Ala 435 440 445 Met Thr Lys Asp Asn Asn Leu Leu Gly Lys Phe Glu Leu Thr Gly Ile 450 455 460 Pro Pro Ala Pro Arg Gly Val Pro Gln Ile Glu Val Thr Phe Asp Ile 465 470 475 480 Asp Ala Asn Gly Ile Leu Asn Val Ser Ala Val Asp Lys Ser Thr Gly 485 490 495 Lys Glu Asn Lys Ile Thr Ile Thr Asn Asp Lys Gly Arg Leu Ser Lys 500 505 510 Glu Asp Ile Glu Arg Met Val Gln Glu Ala Glu Lys Tyr Lys Ala Glu 515 520 525 Asp Glu Lys Gln Arg Asp Lys Val Ser Ser Lys Asn Ser Leu Glu Ser 530 535 540 Tyr Ala Phe Asn Met Lys Ala Thr Val Glu Asp Glu Lys Leu Gln Gly 545 550 555 560 Lys Ile Asn Asp Glu Asp Lys Gln Lys Ile Leu Asp Lys Cys Asn Glu 565 570 575 Ile Ile Asn Trp Leu Asp Lys Asn Gln Thr Ala Glu Lys Glu Glu Phe 580 585 590 Glu His Gln Gln Lys Glu Leu Glu Lys Val Cys Asn Pro Ile Ile Thr 595 600 605 Lys Leu Tyr Gln Ser Ala Gly Gly Met Pro Gly Gly Met Pro Gly Gly 610 615 620 Phe Pro Gly Gly Gly Ala Pro Pro Ser Gly Gly Ala Ser Ser Gly Pro 625 630 635 640 Thr Ile Glu Glu Val Asp 645 75188PRTHomo sapiens 75Met Ala Leu Thr Phe Ala Leu Leu Val Ala Leu Leu Val Leu Ser Cys 1 5 10 15 Lys Ser Ser Cys Ser Val Gly Cys Asp Leu Pro Gln Thr His Ser Leu 20 25 30 Gly Ser Arg Arg Thr Leu Met Leu Leu Ala Gln Met Arg Arg Ile Ser 35 40 45 Leu Phe Ser Cys Leu Lys Asp Arg His Asp Phe Gly Phe Pro Gln Glu 50 55 60 Glu Phe Gly Asn Gln Phe Gln Lys Ala Glu Thr Ile Pro Val Leu His 65 70 75 80 Glu Met Ile Gln Gln Ile Phe Asn Leu Phe Ser Thr Lys Asp Ser Ser 85 90 95 Ala Ala Trp Asp Glu Thr Leu Leu Asp Lys Phe Tyr Thr Glu Leu Tyr 100 105 110 Gln Gln Leu Asn Asp Leu Glu Ala Cys Val Ile Gln Gly Val Gly Val 115 120 125 Thr Glu Thr Pro Leu Met Lys Glu Asp Ser Ile Leu Ala Val Arg Lys 130 135 140 Tyr Phe Gln Arg Ile Thr Leu Tyr Leu Lys Glu Lys Lys Tyr Ser Pro 145 150 155 160 Cys Ala Trp Glu Val Val Arg Ala Glu Ile Met Arg Ser Phe Ser Leu 165 170 175 Ser Thr Asn Leu Gln Glu Ser Leu Arg Ser Lys Glu 180 185 76153PRTHomo sapiens 76Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu 1 5 10 15 Val Thr Asn Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu 20 25 30 Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile 35 40 45 Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe 50 55 60 Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu 65 70 75 80 Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys 85 90 95 Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile 100 105 110 Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala 115 120 125 Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe 130 135 140 Cys Gln Ser Ile Ile Ser Thr Leu Thr 145 150 77407PRTHomo sapiens 77Met Asp Gly Leu Pro Gly Arg Ala Leu Gly Ala Ala Cys Leu Leu Leu 1 5 10 15 Leu Ala Ala Gly Trp Leu Gly Pro Glu Ala Trp Gly Ser Pro Thr Pro 20 25 30 Pro Pro Thr Pro Ala Ala Pro Pro Pro Pro Pro Pro Pro Gly Ser Pro 35 40 45 Gly Gly Ser Gln Asp Thr Cys Thr Ser Cys Gly Gly Phe Arg Arg Pro 50 55 60 Glu Glu Leu Gly Arg Val Asp Gly Asp Phe Leu Glu Ala Val Lys Arg 65 70 75 80 His Ile Leu Ser Arg Leu Gln Met Arg Gly Arg Pro Asn Ile Thr His 85 90 95 Ala Val Pro Lys Ala Ala Met Val Thr Ala Leu Arg Lys Leu His Ala 100 105 110 Gly Lys Val Arg Glu Asp Gly Arg Val Glu Ile Pro His Leu Asp Gly 115 120 125 His Ala Ser Pro Gly Ala Asp Gly Gln Glu Arg Val Ser Glu Ile Ile 130 135 140 Ser Phe Ala Glu Thr Asp Gly Leu Ala Ser Ser Arg Val Arg Leu Tyr 145 150 155 160 Phe Phe Ile Ser Asn Glu Gly Asn Gln Asn Leu Phe Val Val Gln Ala 165 170 175 Ser Leu Trp Leu Tyr Leu Lys Leu Leu Pro Tyr Val Leu Glu Lys Gly 180 185 190 Ser Arg Arg Lys Val Arg Val Lys Val Tyr Phe Gln Glu Gln Gly His 195 200 205 Gly Asp Arg Trp Asn Met Val Glu Lys Arg Val Asp Leu Lys Arg Ser 210 215 220 Gly Trp His Thr Phe Pro Leu Thr Glu Ala Ile Gln Ala Leu Phe Glu 225 230 235 240 Arg Gly Glu Arg Arg Leu Asn Leu Asp Val Gln Cys Asp Ser Cys Gln 245 250 255 Glu Leu Ala Val Val Pro Val Phe Val Asp Pro Gly Glu Glu Ser His 260 265 270 Arg Pro Phe Val Val Val Gln Ala Arg Leu Gly Asp Ser Arg His Arg 275 280 285 Ile Arg Lys Arg Gly Leu Glu Cys Asp Gly Arg Thr Asn Leu Cys Cys 290 295 300 Arg Gln Gln Phe Phe Ile Asp Phe Arg Leu Ile Gly Trp Asn Asp Trp 305 310 315 320 Ile Ile Ala Pro Thr Gly Tyr Tyr Gly Asn Tyr Cys Glu Gly Ser Cys 325 330 335 Pro Ala Tyr Leu Ala Gly Val Pro Gly Ser Ala Ser Ser Phe His Thr 340 345 350 Ala Val Val Asn Gln Tyr Arg Met Arg Gly Leu Asn Pro Gly Thr Val 355 360 365 Asn Ser Cys Cys Ile Pro Thr Lys Leu Ser Thr Met Ser Met Leu Tyr 370 375 380 Phe Asp Asp Glu Tyr Asn Ile Val Lys Arg Asp Val Pro Asn Met Ile 385 390 395 400 Val Glu Glu Cys Gly Cys Ala 405 78582PRTHomo sapiens 78Met Ser Pro Ala Pro Arg Pro Pro Arg Cys Leu Leu Leu Pro Leu Leu 1 5 10 15 Thr Leu Gly Thr Ala Leu Ala Ser Leu Gly Ser Ala Gln Ser Ser Ser 20 25 30 Phe Ser Pro Glu Ala Trp Leu Gln Gln Tyr Gly Tyr Leu Pro Pro Gly 35 40 45 Asp Leu Arg Thr His Thr Gln Arg Ser Pro Gln Ser Leu Ser Ala Ala 50 55 60 Ile Ala Ala Met Gln Lys Phe Tyr Gly Leu Gln Val Thr Gly Lys Ala 65 70 75 80 Asp Ala Asp Thr Met Lys Ala Met Arg Arg Pro Arg Cys Gly Val Pro 85 90 95 Asp Lys Phe Gly Ala Glu Ile Lys Ala Asn Val Arg Arg Lys Arg Tyr 100 105 110 Ala Ile Gln Gly Leu Lys Trp Gln His Asn Glu Ile Thr Phe Cys Ile 115 120 125 Gln Asn Tyr Thr Pro Lys Val Gly Glu Tyr Ala Thr Tyr Glu Ala Ile 130 135 140 Arg Lys Ala Phe Arg Val Trp Glu Ser Ala Thr Pro Leu Arg Phe Arg 145 150 155 160 Glu Val Pro Tyr Ala Tyr Ile Arg Glu Gly His Glu Lys Gln Ala Asp 165 170 175 Ile Met Ile Phe Phe Ala Glu Gly Phe His Gly Asp Ser Thr Pro Phe 180 185 190 Asp Gly Glu Gly Gly Phe Leu Ala His Ala Tyr Phe Pro Gly Pro Asn 195 200 205 Ile Gly Gly Asp Thr His Phe Asp Ser Ala Glu Pro Trp Thr Val Arg 210 215 220 Asn Glu Asp Leu Asn Gly Asn Asp Ile Phe Leu Val Ala Val His Glu 225 230 235 240 Leu Gly His Ala Leu Gly Leu Glu His Ser Ser Asp Pro Ser Ala Ile 245 250 255 Met Ala Pro Phe Tyr Gln Trp Met Asp Thr Glu Asn Phe Val Leu Pro 260 265 270 Asp Asp Asp Arg Arg Gly Ile Gln Gln Leu Tyr Gly Gly Glu Ser Gly 275 280 285 Phe Pro Thr Lys Met Pro Pro Gln Pro Arg Thr Thr Ser Arg Pro Ser 290 295 300 Val Pro Asp Lys Pro Lys Asn Pro Thr Tyr Gly Pro Asn Ile Cys Asp 305 310 315 320 Gly Asn Phe Asp Thr Val Ala Met Leu Arg Gly Glu Met Phe Val Phe 325 330 335 Lys Glu Arg Trp Phe Trp Arg Val Arg Asn Asn Gln Val Met Asp Gly 340 345 350 Tyr Pro Met Pro Ile Gly Gln Phe Trp Arg Gly Leu Pro Ala Ser Ile 355 360 365 Asn Thr Ala Tyr Glu Arg Lys Asp Gly Lys Phe Val Phe Phe Lys Gly 370 375 380 Asp Lys His Trp Val Phe Asp Glu Ala Ser Leu Glu Pro Gly Tyr Pro 385 390 395 400 Lys His Ile Lys Glu Leu Gly Arg Gly Leu Pro Thr Asp Lys Ile Asp 405 410 415 Ala Ala Leu Phe Trp Met Pro Asn Gly Lys Thr Tyr Phe Phe Arg Gly 420 425 430 Asn Lys Tyr Tyr Arg Phe Asn Glu Glu Leu Arg Ala Val Asp Ser Glu 435 440 445 Tyr Pro Lys Asn Ile Lys Val Trp Glu Gly Ile Pro Glu Ser Pro Arg 450 455 460 Gly Ser Phe Met Gly Ser Asp Glu Val Phe Thr Tyr Phe Tyr Lys Gly 465 470 475 480 Asn Lys Tyr Trp Lys Phe Asn Asn Gln Lys Leu Lys Val Glu Pro Gly 485 490 495 Tyr Pro Lys Ser Ala Leu Arg Asp Trp Met Gly Cys Pro Ser Gly Gly 500 505 510 Arg Pro Asp Glu Gly Thr Glu Glu Glu Thr Glu Val Ile Ile Ile Glu 515 520 525 Val Asp Glu Glu Gly Gly Gly Ala Val Ser Ala Ala Ala Val Val Leu 530 535 540 Pro Val Leu Leu Leu Leu Leu Val Leu Ala Val Gly Leu Ala Val Phe 545 550 555 560 Phe Phe Arg Arg His Gly Thr Pro Arg Arg Leu Leu Tyr Cys Gln Arg 565 570 575 Ser Leu

Leu Asp Lys Val 580 79418PRTHomo sapiens 79Met Arg Ala Ala Pro Leu Leu Leu Ala Arg Ala Ala Ser Leu Ser Leu 1 5 10 15 Gly Phe Leu Phe Leu Leu Phe Phe Trp Leu Asp Arg Ser Val Leu Ala 20 25 30 Lys Glu Leu Lys Phe Val Thr Leu Val Phe Arg His Gly Asp Arg Ser 35 40 45 Pro Ile Asp Thr Phe Pro Thr Asp Pro Ile Lys Glu Ser Ser Trp Pro 50 55 60 Gln Gly Phe Gly Gln Leu Thr Gln Leu Gly Met Glu Gln His Tyr Glu 65 70 75 80 Leu Gly Glu Tyr Ile Arg Lys Arg Tyr Arg Lys Phe Leu Asn Glu Ser 85 90 95 Tyr Lys His Glu Gln Val Tyr Ile Arg Ser Thr Asp Val Asp Arg Thr 100 105 110 Leu Met Ser Ala Met Thr Asn Leu Ala Ala Leu Phe Pro Pro Glu Gly 115 120 125 Val Ser Ile Trp Asn Pro Ile Leu Leu Trp Gln Pro Ile Pro Val His 130 135 140 Thr Val Pro Leu Ser Glu Asp Gln Leu Leu Tyr Leu Pro Phe Arg Asn 145 150 155 160 Cys Pro Arg Phe Gln Glu Leu Glu Ser Glu Thr Leu Lys Ser Glu Glu 165 170 175 Phe Gln Lys Arg Leu His Pro Tyr Lys Asp Phe Ile Ala Thr Leu Gly 180 185 190 Lys Leu Ser Gly Leu His Gly Gln Asp Leu Phe Gly Ile Trp Ser Lys 195 200 205 Val Tyr Asp Pro Leu Tyr Cys Glu Ser Val His Asn Phe Thr Leu Pro 210 215 220 Ser Trp Ala Thr Glu Asp Thr Met Thr Lys Leu Arg Glu Leu Ser Glu 225 230 235 240 Leu Ser Leu Leu Ser Leu Tyr Gly Ile His Lys Gln Lys Glu Lys Ser 245 250 255 Arg Leu Gln Gly Gly Val Leu Val Asn Glu Ile Leu Asn His Met Lys 260 265 270 Arg Ala Thr Gln Ile Pro Ser Tyr Lys Lys Leu Ile Met Tyr Ser Ala 275 280 285 His Asp Thr Thr Val Ser Gly Leu Gln Met Ala Leu Asp Val Tyr Asn 290 295 300 Gly Leu Leu Pro Pro Tyr Ala Ser Cys His Leu Thr Glu Leu Tyr Phe 305 310 315 320 Glu Lys Gly Glu Tyr Phe Val Glu Met Tyr Tyr Arg Asn Glu Thr Gln 325 330 335 His Glu Pro Tyr Pro Leu Met Leu Pro Gly Cys Ser Pro Ser Cys Pro 340 345 350 Leu Glu Arg Phe Ala Glu Leu Val Gly Pro Val Ile Pro Gln Asp Trp 355 360 365 Ser Thr Glu Cys Met Thr Thr Asn Ser His Gln Val Leu Lys Val Ile 370 375 380 Phe Ala Val Ala Phe Cys Leu Ile Ser Ala Val Leu Met Val Leu Leu 385 390 395 400 Phe Ile His Ile Arg Arg Gly Leu Cys Trp Gln Arg Glu Ser Tyr Gly 405 410 415 Asn Ile 80386PRTHomo sapiens 80Met Arg Ala Ala Pro Leu Leu Leu Ala Arg Ala Ala Ser Leu Ser Leu 1 5 10 15 Gly Phe Leu Phe Leu Leu Phe Phe Trp Leu Asp Arg Ser Val Leu Ala 20 25 30 Lys Glu Leu Lys Phe Val Thr Leu Val Phe Arg His Gly Asp Arg Ser 35 40 45 Pro Ile Asp Thr Phe Pro Thr Asp Pro Ile Lys Glu Ser Ser Trp Pro 50 55 60 Gln Gly Phe Gly Gln Leu Thr Gln Leu Gly Met Glu Gln His Tyr Glu 65 70 75 80 Leu Gly Glu Tyr Ile Arg Lys Arg Tyr Arg Lys Phe Leu Asn Glu Ser 85 90 95 Tyr Lys His Glu Gln Val Tyr Ile Arg Ser Thr Asp Val Asp Arg Thr 100 105 110 Leu Met Ser Ala Met Thr Asn Leu Ala Ala Leu Phe Pro Pro Glu Gly 115 120 125 Val Ser Ile Trp Asn Pro Ile Leu Leu Trp Gln Pro Ile Pro Val His 130 135 140 Thr Val Pro Leu Ser Glu Asp Gln Leu Leu Tyr Leu Pro Phe Arg Asn 145 150 155 160 Cys Pro Arg Phe Gln Glu Leu Glu Ser Glu Thr Leu Lys Ser Glu Glu 165 170 175 Phe Gln Lys Arg Leu His Pro Tyr Lys Asp Phe Ile Ala Thr Leu Gly 180 185 190 Lys Leu Ser Gly Leu His Gly Gln Asp Leu Phe Gly Ile Trp Ser Lys 195 200 205 Val Tyr Asp Pro Leu Tyr Cys Glu Ser Val His Asn Phe Thr Leu Pro 210 215 220 Ser Trp Ala Thr Glu Asp Thr Met Thr Lys Leu Arg Glu Leu Ser Glu 225 230 235 240 Leu Ser Leu Leu Ser Leu Tyr Gly Ile His Lys Gln Lys Glu Lys Ser 245 250 255 Arg Leu Gln Gly Gly Val Leu Val Asn Glu Ile Leu Asn His Met Lys 260 265 270 Arg Ala Thr Gln Ile Pro Ser Tyr Lys Lys Leu Ile Met Tyr Ser Ala 275 280 285 His Asp Thr Thr Val Ser Gly Leu Gln Met Ala Leu Asp Val Tyr Asn 290 295 300 Gly Leu Leu Pro Pro Tyr Ala Ser Cys His Leu Thr Glu Leu Tyr Phe 305 310 315 320 Glu Lys Gly Glu Tyr Phe Val Glu Met Tyr Tyr Arg Asn Glu Thr Gln 325 330 335 His Glu Pro Tyr Pro Leu Met Leu Pro Gly Cys Ser Pro Ser Cys Pro 340 345 350 Leu Glu Arg Phe Ala Glu Leu Val Gly Pro Val Ile Pro Gln Asp Trp 355 360 365 Ser Thr Glu Cys Met Thr Thr Asn Ser His Gln Gly Thr Glu Asp Ser 370 375 380 Thr Asp 385 81646PRTHomo sapiens 81Met Gly Leu Pro Arg Leu Val Cys Ala Phe Leu Leu Ala Ala Cys Cys 1 5 10 15 Cys Cys Pro Arg Val Ala Gly Val Pro Gly Glu Ala Glu Gln Pro Ala 20 25 30 Pro Glu Leu Val Glu Val Glu Val Gly Ser Thr Ala Leu Leu Lys Cys 35 40 45 Gly Leu Ser Gln Ser Gln Gly Asn Leu Ser His Val Asp Trp Phe Ser 50 55 60 Val His Lys Glu Lys Arg Thr Leu Ile Phe Arg Val Arg Gln Gly Gln 65 70 75 80 Gly Gln Ser Glu Pro Gly Glu Tyr Glu Gln Arg Leu Ser Leu Gln Asp 85 90 95 Arg Gly Ala Thr Leu Ala Leu Thr Gln Val Thr Pro Gln Asp Glu Arg 100 105 110 Ile Phe Leu Cys Gln Gly Lys Arg Pro Arg Ser Gln Glu Tyr Arg Ile 115 120 125 Gln Leu Arg Val Tyr Lys Ala Pro Glu Glu Pro Asn Ile Gln Val Asn 130 135 140 Pro Leu Gly Ile Pro Val Asn Ser Lys Glu Pro Glu Glu Val Ala Thr 145 150 155 160 Cys Val Gly Arg Asn Gly Tyr Pro Ile Pro Gln Val Ile Trp Tyr Lys 165 170 175 Asn Gly Arg Pro Leu Lys Glu Glu Lys Asn Arg Val His Ile Gln Ser 180 185 190 Ser Gln Thr Val Glu Ser Ser Gly Leu Tyr Thr Leu Gln Ser Ile Leu 195 200 205 Lys Ala Gln Leu Val Lys Glu Asp Lys Asp Ala Gln Phe Tyr Cys Glu 210 215 220 Leu Asn Tyr Arg Leu Pro Ser Gly Asn His Met Lys Glu Ser Arg Glu 225 230 235 240 Val Thr Val Pro Val Phe Tyr Pro Thr Glu Lys Val Trp Leu Glu Val 245 250 255 Glu Pro Val Gly Met Leu Lys Glu Gly Asp Arg Val Glu Ile Arg Cys 260 265 270 Leu Ala Asp Gly Asn Pro Pro Pro His Phe Ser Ile Ser Lys Gln Asn 275 280 285 Pro Ser Thr Arg Glu Ala Glu Glu Glu Thr Thr Asn Asp Asn Gly Val 290 295 300 Leu Val Leu Glu Pro Ala Arg Lys Glu His Ser Gly Arg Tyr Glu Cys 305 310 315 320 Gln Gly Leu Asp Leu Asp Thr Met Ile Ser Leu Leu Ser Glu Pro Gln 325 330 335 Glu Leu Leu Val Asn Tyr Val Ser Asp Val Arg Val Ser Pro Ala Ala 340 345 350 Pro Glu Arg Gln Glu Gly Ser Ser Leu Thr Leu Thr Cys Glu Ala Glu 355 360 365 Ser Ser Gln Asp Leu Glu Phe Gln Trp Leu Arg Glu Glu Thr Gly Gln 370 375 380 Val Leu Glu Arg Gly Pro Val Leu Gln Leu His Asp Leu Lys Arg Glu 385 390 395 400 Ala Gly Gly Gly Tyr Arg Cys Val Ala Ser Val Pro Ser Ile Pro Gly 405 410 415 Leu Asn Arg Thr Gln Leu Val Asn Val Ala Ile Phe Gly Pro Pro Trp 420 425 430 Met Ala Phe Lys Glu Arg Lys Val Trp Val Lys Glu Asn Met Val Leu 435 440 445 Asn Leu Ser Cys Glu Ala Ser Gly His Pro Arg Pro Thr Ile Ser Trp 450 455 460 Asn Val Asn Gly Thr Ala Ser Glu Gln Asp Gln Asp Pro Gln Arg Val 465 470 475 480 Leu Ser Thr Leu Asn Val Leu Val Thr Pro Glu Leu Leu Glu Thr Gly 485 490 495 Val Glu Cys Thr Ala Ser Asn Asp Leu Gly Lys Asn Thr Ser Ile Leu 500 505 510 Phe Leu Glu Leu Val Asn Leu Thr Thr Leu Thr Pro Asp Ser Asn Thr 515 520 525 Thr Thr Gly Leu Ser Thr Ser Thr Ala Ser Pro His Thr Arg Ala Asn 530 535 540 Ser Thr Ser Thr Glu Arg Lys Leu Pro Glu Pro Glu Ser Arg Gly Val 545 550 555 560 Val Ile Val Ala Val Ile Val Cys Ile Leu Val Leu Ala Val Leu Gly 565 570 575 Ala Val Leu Tyr Phe Leu Tyr Lys Lys Gly Lys Leu Pro Cys Arg Arg 580 585 590 Ser Gly Lys Gln Glu Ile Thr Leu Pro Pro Ser Arg Lys Ser Glu Leu 595 600 605 Val Val Glu Val Lys Ser Asp Lys Leu Pro Glu Glu Met Gly Leu Leu 610 615 620 Gln Gly Ser Ser Gly Asp Lys Arg Ala Pro Gly Asp Gln Gly Glu Lys 625 630 635 640 Tyr Ile Asp Leu Arg His 645 8220PRTArtificial SequenceSynthetic peptides 82Thr Lys Ala Gly Arg Gly Ala Ser Gln Pro Pro Thr Pro Thr Pro Ala 1 5 10 15 Ser Asp Ala Phe 20 8320PRTArtificial SequenceSynthetic peptides 83Glu Glu Glu Pro Glu Glu Thr Ala Glu Asp Thr Thr Glu Asp Thr Glu 1 5 10 15 Gln Asp Glu Asp 20 8420PRTArtificial SequenceSynthetic peptides 84Thr Ala Ser Thr Thr Ala Asn Thr Pro Phe Pro Thr Ala Thr Ser Pro 1 5 10 15 Ala Pro Pro Ile 20 8520PRTArtificial SequenceSynthetic peptides 85Pro Ala Pro Pro Ile Ile Ser Thr His Ser Ser Ser Thr Ile Pro Thr 1 5 10 15 Pro Ala Pro Pro 20 8620PRTArtificial SequenceSynthetic peptides 86Leu Ala Lys Met Tyr Tyr Ser Ala Val Glu Pro Thr Lys Asp Ile Phe 1 5 10 15 Thr Gly Leu Ile 20 8720PRTArtificial SequenceSynthetic peptides 87Pro Ser Ser Thr Lys Pro Pro Ala Leu Ser His Ser Val Ser Thr Ser 1 5 10 15 Ser Thr Thr Lys 20 8820PRTArtificial SequenceSynthetic peptides 88Leu Glu Pro Asp Tyr Phe Lys Asp Met Thr Pro Thr Ile Arg Lys Thr 1 5 10 15 Gln Lys Ile Val 20 8920PRTArtificial SequenceSynthetic peptides 89Ser Thr Met Pro Val Val Ser Ser Glu Ala Ser Thr His Ser Thr Thr 1 5 10 15 Pro Val Asp Thr 20 9020PRTArtificial SequenceSynthetic peptides 90Ser Thr His Ser Thr Thr Pro Val Asp Thr Ser Thr Pro Val Thr Thr 1 5 10 15 Ser Thr Glu Ala 20 9120PRTArtificial SequenceSynthetic peptides 91Ile Pro Pro Glu Asp Thr Ala Ser Thr Arg Ser Ser Phe Thr Val Gln 1 5 10 15 Asp Leu Lys Pro 20 9220PRTArtificial SequenceSynthetic peptides 92Glu Ala Lys Thr Ser Asn Pro Thr Ser Ser Leu Thr Ser Leu Ser Val 1 5 10 15 Ala Pro Thr Phe 20 9320PRTArtificial SequenceSynthetic peptides 93Ala Arg Thr Glu Pro Trp Glu Gly Asn Ser Ser Thr Ala Ala Thr Thr 1 5 10 15 Pro Glu Thr Phe 20 9420PRTArtificial SequenceSynthetic peptides 94Val Asp Pro Leu Gln Leu Gln Thr Pro Pro Gln Thr Gln Pro Gly Pro 1 5 10 15 Ser His Val Met 20 9520PRTArtificial SequenceSynthetic peptides 95Ser Pro Lys Pro Ser Thr Thr Asn Val Phe Thr Ser Ala Val Asp Gln 1 5 10 15 Thr Ile Thr Pro 20 9620PRTArtificial SequenceSynthetic peptides 96Pro Ala Pro Pro Ala Pro Gly Asn Ala Ser Glu Ser Glu Glu Asp Arg 1 5 10 15 Ser Ala Gly Ser 20 9720PRTArtificial SequenceSynthetic peptides 97Ala Ser Glu Ser Glu Glu Asp Arg Ser Ala Gly Ser Val Glu Ser Pro 1 5 10 15 Ser Val Ser Ser 20 9820PRTArtificial SequenceSynthetic peptides 98Ala Asn Leu Asn Ser Asp Lys Glu Asn Ile Thr Thr Ser Asn Leu Lys 1 5 10 15 Ala Ser His Ser 20 9920PRTArtificial SequenceSynthetic peptides 99Leu Thr Thr Asn Ser Asp Ser Phe Thr Gly Phe Thr Pro Tyr Gln Glu 1 5 10 15 Lys Thr Thr Leu 20 10020PRTArtificial SequenceSynthetic peptides 100Ser Phe Thr Gly Phe Thr Pro Tyr Gln Glu Lys Thr Thr Leu Gln Pro 1 5 10 15 Thr Leu Lys Phe 20 10120PRTArtificial SequenceSynthetic peptides 101Ser Phe Thr Gly Phe Thr Pro Tyr Gln Glu Lys Thr Thr Leu Gln Pro 1 5 10 15 Thr Leu Lys Phe 20 10220PRTArtificial SequenceSynthetic peptides 102Ser Pro Thr Ser Ser Ala Ser Ser Phe Ser Ser Ser Ala Pro Phe Leu 1 5 10 15 Ala Ser Ala Val 20 10320PRTArtificial SequenceSynthetic peptides 103Thr Val Pro Cys Pro Val Pro Ser Thr Pro Pro Thr Pro Ser Pro Ser 1 5 10 15 Thr Pro Pro Thr 20 10420PRTArtificial SequenceSynthetic peptides 104Val Pro Ser Thr Pro Pro Thr Pro Ser Pro Ser Thr Pro Pro Thr Pro 1 5 10 15 Ser Pro Ser Cys 20 10520PRTArtificial SequenceSynthetic peptides 105Ser Glu Pro Pro Lys Ala Ala Arg Pro Pro Val Thr Pro Val Leu Leu 1 5 10 15 Glu Lys Lys Ser 20 10620PRTArtificial SequenceSynthetic peptides 106Gly Gln Ser Gln Pro Thr Val Ala Gly Gln Pro Ser Ala Arg Pro Ala 1 5 10 15 Ala Glu Glu Tyr 20 10720PRTArtificial SequenceSynthetic peptides 107Thr Pro Ala Ser Ile Thr Ala Ala Lys Thr Ser Thr Ile Thr Thr Ala 1 5 10 15 Phe Pro Pro Val 20 10820PRTArtificial SequenceSynthetic peptides 108Gly Asn Ala Ser Met Asp Ala Val Cys Thr Ser Thr Ser Pro Thr Arg 1 5 10 15 Ser Met Ala Pro 20 10920PRTArtificial SequenceSynthetic peptides 109Thr Asp Cys Gly Gly Pro Lys Asp His Pro Leu Thr Cys Asp Asp Pro 1 5 10 15 Arg Phe Gln Ala 20 11021PRTArtificial SequenceSynthetic peptides 110Ala Gln Ala Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser 1 5 10 15 Pro Ser Arg Leu Pro 20 11120PRTArtificial SequenceSynthetic peptides 111Pro Leu Ser Glu Leu Glu Ser Gly Glu Gln Pro Ser Asp Glu Gln Pro 1 5 10 15 Ser Gly Glu His 20 11220PRTArtificial SequenceSynthetic peptides 112Pro Gln Arg Ser Ser Thr Ala Ile Leu Gln Val Ser Val Thr Asp Thr 1 5

10 15 Asn Asp Asn His 20 11320PRTArtificial SequenceSynthetic peptides 113Gly Ala Leu Pro Gly Thr Ser Val Met Glu Val Thr Ala Thr Asp Ala 1 5 10 15 Asp Asp Asp Val 20 11420PRTArtificial SequenceSynthetic peptides 114Glu Gln Glu Pro Pro Ser Thr Asp Val Pro Pro Ser Pro Glu Ala Gly 1 5 10 15 Gly Thr Thr Gly 20 11520PRTArtificial SequenceSynthetic peptides 115Arg Pro Glu Ala Thr Pro Phe Leu Val Ala His Thr Arg Thr Arg Pro 1 5 10 15 Pro Ser Gly Gly 20 11620PRTArtificial SequenceSynthetic peptides 116Pro Gly Thr Ser Thr Thr Pro Ser Gln Pro Asn Ser Ala Gly Val Gln 1 5 10 15 Asp Thr Glu Met 20 11720PRTArtificial SequenceSynthetic peptides 117Glu Val Ala Pro Glu Ala Ser Thr Ser Ser Ala Ser Gln Val Ile Ala 1 5 10 15 Pro Thr Gln Val 20 11820PRTArtificial SequenceSynthetic peptides 118Gln Pro Pro Asp Phe Ala Leu Ala Tyr Arg Pro Ser Phe Pro Glu Asp 1 5 10 15 Arg Glu Pro Gln 20 11920PRTArtificial SequenceSynthetic peptides 119Leu Ser Val Thr Arg Pro Val Val Val Ser Ala Thr His Pro Thr Leu 1 5 10 15 Pro Ser Ala His 20 12020PRTArtificial SequenceSynthetic peptides 120Pro Ser Ala His Gln Pro Pro Val Ile Pro Ala Thr His Pro Ala Leu 1 5 10 15 Ser Arg Asp His 20 12120PRTArtificial SequenceSynthetic peptides 121Ala Pro Asp Ala Leu Val Leu Arg Thr Gln Ala Thr Gln Leu Pro Ile 1 5 10 15 Ile Pro Thr Ala 20 12220PRTArtificial SequenceSynthetic peptides 122Gly Ala Leu Phe Pro Gly Pro Gly Asn Ala Gln Thr Ser Val Ser Pro 1 5 10 15 Ser Lys Val Ile 20 12320PRTArtificial SequenceSynthetic peptides 123His Leu Ala Leu Gly Asp Gln Arg Leu Asn Pro Thr Val Thr Tyr Gly 1 5 10 15 Asn Asp Ser Phe 20 12420PRTArtificial SequenceSynthetic peptides 124His Leu Ala Leu Gly Asp Gln Arg Leu Asn Pro Thr Val Thr Tyr Gly 1 5 10 15 Asn Asp Ser Phe 20 12520PRTArtificial SequenceSynthetic peptides 125Pro Ala Thr Glu Pro Ala Ser Gly Ser Ala Ala Thr Trp Gly Gln Asp 1 5 10 15 Val Thr Ser Val 20 12620PRTArtificial SequenceSynthetic peptides 126Val Pro Val Thr Arg Pro Ala Leu Gly Ser Thr Thr Pro Pro Ala His 1 5 10 15 Asp Val Thr Ser 20 12720PRTArtificial SequenceSynthetic peptides 127Asn Val Thr Ser Ala Ser Gly Ser Ala Ser Gly Ser Ala Ser Thr Leu 1 5 10 15 Val His Asn Gly 20 12820PRTArtificial SequenceSynthetic peptides 128Asn Val Thr Ser Ala Ser Gly Ser Ala Ser Gly Ser Ala Ser Thr Leu 1 5 10 15 Val His Asn Gly 20 12920PRTArtificial SequenceSynthetic peptides 129Gly Thr Ser Ala Arg Ala Thr Thr Thr Pro Ala Ser Lys Ser Thr Pro 1 5 10 15 Phe Ser Ile Pro 20 13020PRTArtificial SequenceSynthetic peptides 130Gly Thr Ser Ala Arg Ala Thr Thr Thr Pro Ala Ser Lys Ser Thr Pro 1 5 10 15 Phe Ser Ile Pro 20 13120PRTArtificial SequenceSynthetic peptides 131Ser Asp Thr Pro Thr Thr Leu Ala Ser His Ser Thr Lys Thr Asp Ala 1 5 10 15 Ser Ser Thr His 20 13220PRTArtificial SequenceSynthetic peptides 132Thr Lys Thr Asp Ala Ser Ser Thr His His Ser Thr Val Pro Pro Leu 1 5 10 15 Thr Ser Ser Asn 20 13320PRTArtificial SequenceSynthetic peptides 133Thr Asp Tyr Tyr Gln Glu Leu Gln Arg Asp Ile Ser Glu Met Phe Leu 1 5 10 15 Gln Ile Tyr Lys 20 13420PRTArtificial SequenceSynthetic peptides 134His Asp Val Glu Thr Gln Phe Asn Gln Tyr Lys Thr Glu Ala Ala Ser 1 5 10 15 Arg Tyr Asn Leu 20 13520PRTArtificial SequenceSynthetic peptides 135Ala Ser Arg Tyr Asn Leu Thr Ile Ser Asp Val Ser Val Ser Asp Val 1 5 10 15 Pro Phe Pro Phe 20 13620PRTArtificial SequenceSynthetic peptides 136Arg Tyr Asn Leu Thr Ile Ser Asp Val Ser Val Ser Asp Val Pro Phe 1 5 10 15 Pro Phe Ser Ala 20 13720PRTArtificial SequenceSynthetic peptides 137Asp Val Ser Val Ser Asp Val Pro Phe Pro Phe Ser Ala Gln Ser Gly 1 5 10 15 Ala Gly Val Pro 20 13820PRTArtificial SequenceSynthetic peptides 138Thr Ala Gly Arg Pro Thr Gly Gln Ser Ser Pro Thr Ser Pro Ser Ala 1 5 10 15 Ser Pro Gln Glu 20 13920PRTArtificial SequenceSynthetic peptides 139Ser Leu Ala Ser Gln Ala Thr Asp Thr Phe Ser Thr Val Pro Pro Thr 1 5 10 15 Pro Pro Ser Ile 20 14020PRTArtificial SequenceSynthetic peptides 140Phe Ser Thr Val Pro Pro Thr Pro Pro Ser Ile Thr Ser Thr Gly Leu 1 5 10 15 Thr Ser Pro Gln 20 14120PRTArtificial SequenceSynthetic peptides 141Pro Thr Pro Pro Ser Ile Thr Ser Thr Gly Leu Thr Ser Pro Gln Thr 1 5 10 15 Glu Thr His Thr 20 14220PRTArtificial SequenceSynthetic peptides 142Leu Thr Ser Pro Gln Thr Glu Thr His Thr Leu Ser Pro Ser Gly Ser 1 5 10 15 Gly Lys Thr Phe 20 14320PRTArtificial SequenceSynthetic peptides 143Thr Asp Thr Ser Ser Ala Ser Thr Gly His Ala Thr Pro Leu Pro Val 1 5 10 15 Thr Ser Leu Ser 20 14420PRTArtificial SequenceSynthetic peptides 144His Ala Thr Pro Leu Ala Val Ser Ser Ala Thr Ser Ala Ser Thr Val 1 5 10 15 Ser Ser Asp Ser 20 14520PRTArtificial SequenceSynthetic peptides 145Val Pro Val Thr Arg Pro Ala Leu Gly Ser Thr Thr Pro Pro Ala His 1 5 10 15 Asp Val Thr Ser 20 14620PRTArtificial SequenceSynthetic peptides 146Ser Leu Ala Ser Gln Ala Thr Asp Thr Phe Ser Thr Val Pro Pro Thr 1 5 10 15 Pro Pro Ser Ile 20

Claims

1. A method for identifying glycopeptides reactive with cancer-associated auto-antibodies, the method comprising: providing a panel comprising peptides immobilized on said panel, at least a plurality of said peptides comprising one or more sites amenable to glycosylation, wherein at least one of said glycosylation sites is modified with a glycan to form a glycopeptide having a peptide portion and a glycan portion; contacting said panel with an antibody-containing sample from a patient with cancer; and identifying glycopeptides in said panel that (i) are selectively recognized by antibodies in said sample, but not by antibodies in a control sample and (ii) are recognized by such antibodies in said sample through recognition of both the peptide portion and the glycan portion and not through recognition of either the peptide or the glycan alone.

2. The method of claim 1, wherein said panel comprises peptides having an amino acid sequence comprising at least one serine or threonine residue, wherein said residue is a glycosylation site.

3. The method of claim 1, wherein said control sample contains pooled samples from a plurality of control individuals.

4. The method of claim 1, wherein said panel comprises mutants of said peptides.

5. The method of claim 1, further comprising identifying minimum cancer-associated glycopeptide epitopes.

6. The method of claim 5, further comprising elucidating the epitope structure of said identified glycopeptide epitopes.

7. The method of claim 1, wherein each of said peptides in said panel is about 2 to about 50 amino acid residues in length.

8. The method of claim 7, wherein each of said peptides in said panel is about 4 to about 25 amino acid residues in length.

9. The method of claim 1, wherein said amino acid sequence of said peptides is a fragment found in a protein or variant of said protein or a conservative mutant.

10. The method of claim 1, wherein said identified glycopeptides are found in at least one glycoprotein that is aberrantly glycosylated in cancer cells.

11. The method of claim 1, wherein said identified glycopeptides are found in at least one glycoprotein that is overexpressed in cancer cells.

12. The method of claim 1, wherein the serine or threonine residue is placed at about the center of each of said amino acid sequences.

13. The method of claim 1, wherein said glycopeptides are synthesized synthetically or chemoenzymatically.

14. The method of claim 1, wherein said glycopeptides of said panel are partially glycosylated peptides.

15. The method of claim 1 or 14, wherein said glycopeptides of said panel are glycosylated in situ, in solution, or in vivo by recombinant expression in a host cell.

16. The method of claim 1, wherein said glycopeptides of said panel are treated with one or more exoglycosidases to expose O-glycans.

17. The method of claim 1, wherein said glycan is an O-glycan.

18. The method of claim 17, wherein said O-glycan is a member selected from the group consisting of: Tn, STn, T, Truncated C3, Truncated C2, Truncated C4, non-capped type1-C3, non-capped type2-C2, non-capped type2-C4, GalNAca-3Tn, SA-type1-C3, SLea-C3, LacDiNAc-C3, LacDiNAc-C2, and LacDiNAc-C4.

19. The method of claim 1, wherein said panel is a microarray.

20. The method of claim 1, wherein said identified glycopeptide is selectively recognized by a cancer-associated IgG antibody.

21. The method of claim 1, wherein said panel comprises one or more glycopeptides having an amino acid sequence selected from the group consisting of: SHHSDESDELVTDFPTDLPA (SEQ ID NO: 15); TPTPKEKPEAGTYSVNNGND (SEQ ID NO: 36); SESFPHPGFNMSLLENHTRQ (SEQ ID NO: 49); LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTCDDPRFQA (SEQ ID NO: 109); PGTSTTPSQPNSAGVQDTEM (SEQ ID NO: 116); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132); HDVETQFNQYKTEAASRYNL (SEQ ID NO: 134); ASRYNLTISDVSVSDVPFPF (SEQ ID NO: 135); VPVTRPALGSTTPPAHDVTS (SEQ ID NO: 145); and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

22. A pharmaceutical composition for eliciting an immune response in a patient, comprising one or more isolated glycopeptides consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 86, 109, 116, 132, 134, 135 and 146, and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition of claim 22, wherein the one or more isolated glycopeptides are selected from the group consisting of LAKMYYSAVEPTKDIFTGLI (SEQ ID NO: 86); TDCGGPKDHPLTDPRFQA (SEQ ID NO: 109); TKTDASSTHHSTVPPLTSSN (SEQ ID NO: 132), and SLASQATDTFSTVPPTPPSI (SEQ ID NO: 146).

24. A pharmaceutical composition for eliciting an immune response in a patient, comprising one or more isolated glycopeptides consisting of an epitope of 5 to 10 amino acid residues contained in a glycopeptide selected from the group consisting of SEQ ID NOs: 86, 109, 116, 132, 134, 135 and 146, and a pharmaceutically acceptable carrier, said epitope having been previously determined (i) to be selectively recognized by a subset of antibodies in sera from cancer patients, which subset recognizes neither (a) the corresponding naked peptides of said panel when not glycosylated; nor (b) the corresponding glycan when not bound to said peptide; and (ii) not to be recognized by antibodies in control sera.

25. A panel of glycopeptides comprising at least a plurality of isolated glycopeptides immobilized on said panel, each isolated glycopeptide comprising a glycopeptide epitope, said epitope having been previously determined (i) to be selectively recognized by a subset of antibodies in sera from cancer patients, which subset recognizes neither (a) the corresponding naked peptides of said panel when not glycosylated; nor (b) the corresponding glycan when not bound to said peptide; and (ii) not to be recognized by antibodies in control sera, said plurality comprising at least 8 isolated glycopeptides, wherein each epitope is 5 to 10 amino acid residues contained in one of the amino acid sequences of SEQ ID NOs: 86, 109, 116, 132, 134, 135, 145, and 146, respectively.

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