Abstract:
DNA which encodes a protein homologous to the protein tyrosine phosphatases (PTPases) which catalyze the dephosphorylation of proteins in which tyrosyl residues have been phosphorylated through the action of a protein kinase. The protein, which appears as if it will localize to focal adhesions, is also the subject of the present invention. In particular, it relates to cDNA encoding a protein, referred to as PTPH 1.    
     Overexpression of PTPH 1  may be a powerful approach to countering the effects of oncogenic protein tyrosine kinases, such as those of transforming viruses, and interfering with or reversing cell transformation. This would provide a means of preventing or reversing abnormally high levels of phosphotyrosine associated with any disease or condition, such as preventing or reversing malignancy associated with the activity of a protein tyrosine kinase.  
     In the present method, DNA or RNA encoding PTPH 1  or a functional equivalent of PTPH 1  is administered to an individual in an appropriate gene transfer vehicle which can infect mammalian cells and, once inside the mammalian cells, express and make available PTPH 1  or its functional equivalent in sufficient quantities to overcome or counteract the protein tyrosine kinase activity. As a result, phosphorylation of tyrosine residues at abnormal levels is prevented or reversed, resulting in turn in prevention or reversal of malignancy of cells.

Description:
RELATED APPLICATION  
       [0001]    This application is a continuation of U.S. application Ser. No. 09/235,251, filed Jan. 22, 1999, which is a continuation of U.S. application Ser. No. 08/759,536, filed Dec. 4, 1996, now issued U.S. Pat. No. 5,863,781, which is a divisional of U.S. application Ser. No. 08/107,420, filed Aug. 16, 1993, now issued, U.S. Pat. No. 5,595,911, which is a file wrapper continuation of U.S. application Ser. No. 07/663,579, filed Mar. 1, 1991, now abandoned, which is a continuation-in-part of U.S. application Ser. No. 07/494,036, filed Mar. 14, 1990, now abandoned. The contents of all the above applications are incorporated herein by their entirety. 
     
    
     
       FUNDING  
         [0002]    Work described herein was funded by Cold Spring Harbor Laboratory.  
         BACKGROUND  
         [0003]    Focal adhesion plaques are specialized regions of the plasma membrane through which cells in culture adhere to the external substrate (Burridge, K. et al.,  Ann Rev. Cell. Biol.,  4:487-525 (1988); Burridge, K. and K. Fath,  BioEssays,  10:104-108 (1989)). On their internal face these structures anchor actin stress fibers, which are important in determining cell shape. Similar, but less well-characterized structures have been implicated in attachment between neighboring cells and adherence to extracellular matrix in vivo. Oncogenic transformation is frequently accompanied by a less-adherent, rounded morphology resulting from reorganization of the cytoskeleton (Ben Ze&#39;ev, A.,  Biochem. Biophys. Acta.,  780:197-212 (1985; Felice, G. R. et al.,  Eur. J. Cell Biol.,  52:47-49 (1990)). In Rous Sarcoma Virus (RSV) transformed cells, for instance, it has been postulated that a contributing factor is the aberrant phosphorylation by pp60 v-src  of tyrosyl residues in key focal adhesion proteins (Burr, J. G. et al.,  Proc. Natl. Acad. Sci. USA,  77:3484-3488 (1980); Parsons, J. T. and M. J. Weber,  Curr. Topics in Microbiol. and Immunol.,  147:79-127 (1989)). Phosphotyrosine has also been detected in focal adhesions (Maher, P. A. et al.,  Proc. Natl. Acad. Sci. USA,  82:6576-6580 (1985)) and apical junctions (Takata, K. and S. J. Singer,  J. Cell Biol.,  106:1757-1764 (1988)) in nontransformed cells, raising the possibility that tyrosine phosphorylation at these sites may regulate normal cellular function. Such phosphorylation events must be tightly controlled and an understanding of the mechanism(s) involved would be very useful in furthering our understanding of control of normal and neoplastic cell growth.  
         SUMMARY OF THE INVENTION  
         [0004]    The present invention relates to DNA which encodes a protein homologous to the protein tyrosine phosphatases (PTPases) which catalyze the dephosphorylation of proteins in which tyrosyl residues have been phosphorylated through the action of a protein kinase. The protein, which appears as if it will localize to focal adhesions, is also the subject of the present invention. In particular, it relates to cDNA encoding a protein, referred to as PTPH 1 , which was obtained from HeLa cells and characterized. PTPH 1  has also been identified in other cell types. The structure of PTPH 1  includes three segments: 1) an N-terminal segment of approximately 320 residues, which shows homology with the N-terminal segments of the talin family in the region known to be important for localization to focal adhesions; 2) a central segment, in which there are sequences with the features of sites of phosphorylation by casein kinase 2 and p34 cdc2 , which may be important for regulation of phosphatase activity; and 3) a C-terminal segment of approximately 250 residues, which shows homology to the known members of the PTPase family. PTPH 1  has a single putative catalytic domain.  
           [0005]    Because of its homology with the talin family of proteins, which are known to participate in linkage of intracellular actin filaments to the extracellular matrix at focal adhesions, it is likely that PTPH 1  localizes to the focal adhesions, which is a major site of action for oncogenic protein tyrosine kinases (PTK). Thus, overexpression of PTPH 1  may be a powerful approach to countering the effects of oncogenic protein tyrosine kinases, such as those of transforming viruses, and interfering with or reversing cell transformation. This would provide a means of preventing or reversing abnormally high levels of phosphotyrosine associated with any disease or condition, such as preventing or reversing malignancy associated with the activity of a protein tyrosine kinase. Such protein tyrosine kinase may be of viral origin or be a cellular protein tyrosine kinase whose normal cellular function is disrupted, resulting in abnormal phosphorylation of tyrosyl residues. Such a method of preventing or reversing malignancy caused by or associated with the activity of a protein tyrosine kinase is also the subject of the present invention. In the present method, DNA or RNA encoding PTPH 1  or a functional equivalent of PTPH 1  is administered to an individual in an appropriate gene transfer vehicle which can infect mammalian cells and, once inside the mammalian cells, express and make available PTPH 1  or its functional equivalent in sufficient quantities to overcome or counteract the protein tyrosine kinase activity. As a result, phosphorylation of tyrosine residues at abnormal levels is prevented or reversed, resulting in turn in prevention or reversal of malignancy of cells. Suitable gene transfer vehicles are those which contain DNA or RNA encoding PTPH 1  or a PTPH 1  functional equivalent, can infect mammalian cells and express the encoded protein within the infected mammalian cells. Such vehicles include recombinant retroviruses and recombinant vaccinia virus.  
           [0006]    The method of the present invention is useful in treating or preventing a wide variety of conditions in which abnormally high levels of phosphotyrosine occur and particularly in treating or preventing malignancies in which tyrosyl phosphorylation by a protein tyrosine kinase occurs at an abnormal rate or level and in which dephosphorylation of tyrosyl residues by PTPH 1  or its functional equivalent results in prevention or reversal of a malignant phenotype. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0007]    FIGS.  1 A- 1 B shows the nucleotide sequence and the deduced amino acid sequence of PTPH 1 .  
         [0008]    [0008]FIG. 2 shows the results of northern blot analysis of PTPH 1  mRNA from HeLa cells.  
         [0009]    [0009]FIG. 3 is a schematic diagram illustrating the structure of PTPH 1  and its relationship to CD45, T-cell PTPase and PTPase 1B and human band 4.1, human ezrin and mouse talin.  
         [0010]    [0010]FIG. 4 shows the alignment of the amino acid residues of PTPH 1  and the conserved domains in related proteins. FIG. 4A is a comparison of the sequence of the N-terminal segment PTPH 1  with the homologous domain in band 4.1, ezrin and talin. FIG. 4B is a comparison of the PTPase-like domain in PTPH 1  with the catalytic domains of two low Mr. PTPases (PTP 1 B and TCPTP) and a receptor linked form (CD45). 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0011]    The present invention relates to DNA encoding a protein tyrosine phosphatase which appears to localize to focal adhesions. In particular, it relates to PTPH 1 , which in one segment is homologous to the protein tyrosine phosphatases (PTPases) and in another segment is homologous to a family of proteins which are located at the interface between the plasma membrane and the cytoskeleton. The present invention also relates to the encoded PTPH 1  protein, RNA encoding the PTPH 1  protein, antibodies specific for PTPH 1  protein and methods of using the DNA encoding PTPH 1  and the PTPH 1  protein. DNA encoding PTPH 1  can be used to prevent or reverse phosphorylation of tyrosine residues in proteins which have been phosphorylated by a protein tyrosine kinase.  
         [0012]    As described herein, a protein which is a homologue of the protein tyrosine phosphatases has been identified by means of amplification of PTPase-related cDNAs. In addition, the deduced amino acid sequence of PTPH 1  has been compared with the amino acid sequences of other proteins with known activities. As a result, PTPH 1  has been shown to have three segments. As described below, the characteristics of at least two of these regions make it reasonable to expect that PTPH 1  localizes to the focal adhesions and has intrinsic phosphatase activity.  
         [0013]    The following is a summary of aspects of cytoskeletal components and maintenance of cytoskeletal integrity and description of the identification of PTPH 1 . Also described are the possible role of PTPH 1  in maintaining cytoskeletal integrity and preventing or reversing tyrosyl phosphorylation and a method of preventing or reversing malignant transformation of mammalian cells. In the method of preventing or reversing malignant transformation, DNA or RNA encoding PTPH 1  or a PTPH 1  functional equivalent is introduced into cells and expressed in sufficient quantities to prevent or reverse abnormal tyrosyl phosphorylation, resulting in prevention of transformation of the cells or reversal of the malignant phenotype.  
         [0014]    The actin stress fibers are a component of the cytoskeleton that are important for the determination of cell shape and also for attachment to the substratum. These actin cables interact with the cell membrane at regions described as focal contacts, focal adhesions or adhesion plaques. Fibronectin is an extracellular glycoprotein that is arranged in immobilized fibrillar arrays across the surface of many cells, forming network that interconnects cells with each other. The receptors for fibronectin on the cell surface are termed integrins. Integrins are also localized to focal adhesions and provide the transmembrane link between components of the extracellular matrix, such as fibronectin and vitronectin, and the cytoskeleton.  
         [0015]    Progress has been made in identifying the components of the focal adhesions that provide the link between the integrins and the actin fibers; however, the picture is not yet complete. At present it is thought that the integrins interact indirectly with actin cables through a multiprotein complex. Talin interacts with the cytoplasmic segment of the integrin β chain and also binds to vinculin. Vinculin has been shown to interact with α actinin, which can bind actin directly. Of these components, vinculin, talin and the β subunit of the integrins are phosphorylated on tyrosyl residues. A correlation has yet to be established between such phosphorylation and the appearance of the transformed phenotype. However, this picture is undoubtedly imcomplete. Other substrates of tyrosine phosphorylation, including two focal adhesion associated proteins, paxillin and ezrin, have also been identified. Furthermore, microtubules and intermediate filaments may also terminate at focal adhesions but the nature of the proteins that facilitate this interaction is unknown. Thus, additional focal adhesion associated proteins remain to be characterized and their function established. Protein and cDNA sequence analysis has defined a family of proteins that include talin (Rees, D. J. G. et al.,  Nature,  347:685-689 (1990)), ezrin (Turunen, O. et al.,  J. Biol. Chem.,  264:16727-16732 (1989); Gould, K. L. et al.,  EMBO J.,  8:4133-4142 (1989)) and band 4.1, (Conboy, J. et al.,  Proc. Natl. Acad. Sci. USA,  83:9512-9516 (1986)) which participate in the interaction between the membrane and cytoskeleton. They possess a homologous N-terminal domain that appears to associate with protein components in the plasma membrane. In band 4.1, which promotes association of actin and spectrin in erythrocytes, this domain interacts with the transmembrane protein glycophorin (Bennett, V.,  Biochem. Biophys. Acta.,  988:107-121 (1989)). A similar model has been proposed for the interaction of talin with integrin (Rees, D. J. G. et al.,  Nature,  347:685-689 (1990)). In the case of ezrin, which displays a submembranous localization in brush border cells, the details of its interaction with other proteins remain to be established.  
         [0016]    Identification of the complement of protein tyrosine phosphatases in HeLa cells was carried out as described in detail in Example 1. Briefly, protein tyrosine phosphatase-related cDNAs were amplified in the polymerase chain reaction (PCR) using primers corresponding to conserved segments within the catalytic domains. The PCR reaction products (approximately 0.25 kb) were subcloned into pUC118; of 77 independent subclones sequenced, 15 corresponded to a novel PTPase isoform termed PTPH 1 . A total of 4.2×10 6  phage plaques from the same HeLa cell cDNA library were probed with the PCR product and 10 positive clones were identified.  
         [0017]    The sequence of the longest clone (3984 bp) is presented in FIG. 1. It has a 5′-noncoding segment of 23 bp followed by an open reading frame, from the first available ATG, of 2739 bp. This open reading frame would encode a protein of 913 amino acids with a predicted Mr of approximately 104 k. Although the sequence surrounding the putative initiator does not conform well to the Kozak consensus sequence (Kozak, M.,  J. Cell. Biol.,  108:229-241 (1989)), there is a purine at position −3 which is an important requirement for an initiation site. Furthermore, translation of RNA, synthesized from PTPH 1  cDNA, in a reticulocyte lysate yielded a protein of approximately 120 kDa, close to the expected size. There is a 3′ non-translated segment of 1222 bp, but the cDNA does not contain a consensus polyadenylation signal or a poly A tail. A transcript encoding PTPH 1  was detected by northern blot analysis of HeLa cell polyA+ mRNA as a low abundance message of approximately 4.3 kb (FIG. 2). The slightly larger size of the mRNA may be explained in part by the absence of a poly A tail in the cDNA.  
         [0018]    The sequence of PTPH 1  can be described in terms of 3 segments, as summarized in FIG. 3. The alignment of amino acid residues of PTPH 1  and the conserved domains in related proteins was also assessed. The results are represented in FIG. 4. In FIG. 4A, the sequence of the N-terminal segment of PTPH 1  is compared with the homologous domain in 4.1, ezrin and talin. As shown in FIG. 4A, the N-terminal segment of PTPH 1  is homologous to the domains in band 4.1, ezrin and talin that have been proposed to function in localizing these proteins to the interface between the plasma membrane and the cytoskeleton (Rees, D. J. G. et al.,  Nature,  347:685-689 (1990)).  
         [0019]    In FIG. 4B, the sequence of the protein tyrosine phosphatase-related domain of PTPH 1  is compared with the catalytic domains of two low Mr PTPases (PTP 1 B and TCPTP) and a receptor-linked form (CD45). The C-terminal segment displays homology to the catalytic domain of the PTPases (FIG. 4B). As shown in FIG. 1, the central segment contains sequences that suggest potential phosphorylation sites for casein kinase II and p34 cdc2 . While the dephosphorylation of tyrosyl residues in proteins by PTPH 1  remains to be demonstrated, it should be noted that in CD45 (Tonks, N. K., et al.,  Biochemistry,  27:8695-8701 (1988))) and LAR (Sreuli, M et al.,  EMBO J.,  9:2399-2407 (1990)), which share comparable levels of sequence identity to PTP 1 B as displayed by PTPH 1 , intrinsic activity has been confirmed.  
         [0020]    The PTPases comprise a rapidly expanding family of enzymes and it is anticipated that the various isoforms will have specific functions in vivo. The structure of PTPH 1  may illustrate a general theme among the PTPases; within the protein, distinct structural motifs may, at least in part, control specificity by restricting intracellular localization. It is proposed that PTPH 1  acts at the junction between the cytoskeleton and the plasma membrane and plays a role in controlling cytoskeletal integrity. The occurrence of a signal peptide sequence and a transmembrane domain in CD45 directs it to span the membrane. The C-terminal, non-catalytic segment of the low Mr, cytoplasmic PTPases also appears to direct association with the particulate fraction of cell extracts. In addition such structural motifs may determine how the activity of the catalytic domain is controlled. Thus for the receptor-like forms the binding of ligands to the extracellular segments may modulate activity. The C-terminal segment of the low Mr PTPase appears to repress the activity of the catalytic domain. If PTPH 1  is localized to focal adhesions it should also be regulated to permit normal tyrosine phosphorylation at such sites in non-transformed cells. Phosphorylation of Ser/Thr residues in the central segment of the protein (FIG. 1) may directly modulate activity. In addition, by analogy with band 4.1 whose affinity for glycophorin is regulated by phosphatidylinositol-4,5-bisphosphate, it is possible that the localization of PTPH 1  may alter with phosphatidylinositol turnover.  
         [0021]    Considering the apparent correlation between RSV induced transformation and the cytoskeletal association of pp60 src  (Burr, J. G. et al.,  Proc. Natl. acad. Sci. USA,  77:3484-3488 (1980); Shriver, K. and L. R. Rohrschneider,  Cold Spring Harbor Conference Cell Proliferation,  8: Protein Phosphorylation,  1247-1262 (1981); Hamaguchi, M. and Hanafusa, H.,  Proc. Natl. Acad. Sci. USA,  84:2312-2316 (1987); Tapley, P. et al.,  Oncogene,  4:325-333 (1989); Glenney, J. R., Jr. and Zokas, L.,  J. Cell Biol.,  108:2401-2408 (1989) PTPH 1  is an excellent candidate PTPase with which to achieve a reversion of src-induced transformation and furthermore to delineate the precise role of tyrosine phosphorylation in the morphological changes induced by src. In addition, it clearly has potential to function as growth suppressor; the inactivation or deletion of the PTPH 1  could conceivably in itself be sufficient to generate a transformed phenotype.  
         [0022]    It has been known for some time that viral transformation, for instance by Rous Sarcoma Virus (RSV), brings about a decrease in cell adhesion that is associated with a rounding up of the cells, a reduction in the number of stress fibers and a loss of fibronectin-integrin association. In RSV transformed cells the number of focal adhesions is dramatically decreased and those that remain are altered in architecture. The transforming protein tyrosine kinase of RSV, pp60 v-src , is localized to the adhesion plaques; this is associated with an increased level of phosphotyrosine in these structures. In fact, the level of phosphotyrosine in the focal adhesions correlates with the density of actin stress fibers. That is, the higher the level of phosphotyrosine, the less intact is the cytoskeleton. Therefore, it appears that the association of pp60 v-src  with the focal adhesions and the phosphorylation of tyrosyl residues in proteins within this structure are intimately involved in the disruption of the cytoskeleton.  
         [0023]    Transforming genes, oncogenes, are mutated forms of normal cellular genes, termed proto-oncogenes. Malignant transformation can be described as a disruption of the normal modes of growth control, that is, transformed cells can divide in the absence of specific growth stimulatory factors or fail to respond to growth inhibitory signals. In other words, transformation may often result from the disruption of a normal cellular process. In this regard, it is interesting to note that in the normal process of mitosis, the cells round up and there is a transient disruption of focal contacts.  
         [0024]    Antibodies to phosphotyrosine have indicated that tyrosine phosphorylation of focal adhesion (Maher, P. A. et al.,  Proc. Natl. Acad. Sci. USA,  82:6576-6580 (1985)) and apical junction (Takata, K. and S. J. Singer,  J. Cell Biol.,  106:1757-1764 (1988)) proteins also occurs in non-transformed cells, suggesting the action of a PTK at these sites in normal cell function. However, the kinases involved have yet to be identified. In view of the potentially disastrous effects of aberrant tyrosine phosphorylation one would anticipate that the action of such PTKs would be tightly controlled. The localization of a PTPase to these structures is one means by which such regulation could be achieved.  
         [0025]    The protein tyrosine phosphatases have been shown to constitute a family of both cytoplasmic, low-Mr and trans-membrane receptor-linked forms. The work described herein has resulted in identification of a novel homologue of the protein tyrosine phosphatases in epithelial cells, specifically HeLa cells, which are a model system for the study of the cell cycle. As described, this protein tyrosine phosphatase (PTPH 1 ) has homology with the talin family of proteins and, therefore, can be expected to localize to the interface between the plasma membrane and the cytoskeleton, such as at focal adhesions, which are a major site of action of oncogenic protein tyrosine kinases. Such a protein tyrosine phosphatase is referred to herein as one which localizes to the interface between the plasma membrane and the cytoskeleton. Overexpression of PTPH 1  can be used to counter the effects of protein tyrosine kinases, such as v-src.  
         [0026]    As a result of the findings described herein, a protein tyrosine phosphatase which appears to localize to focal adhesions, which is a major site of action for oncogenic protein tyrosine kinases is available, as are DNA and RNA encoding the protein. The ability of PTPH 1  or a functional equivalent to reverse malignant transformation of cells can be assessed using known methods. For example, this can be carried out as follows: briefly, cells, for instance chicken embryo fibroblasts or NIH 3T3 cells, which are transformed by introduction of a viral oncogene such as v-src, display a change in morphology relative to the non-transformed controls. They will grow in soft agar and induce tumor formation in nude mice. Transformed cells are transfected with the PTPH 1  DNA using an appropriate expression vector. The ability of PTPH 1  DNA or its functional equivalent to have the desired effect is assessed by determining whether the transformed phenotype is maintained or reversed after PTPH 1  DNA or its functional equivalent is introduced into the transformed cells. Reversal of the phenotype is indicative of the ability of the introduced DNA to counter the effects of the kinase. These effects can also be examined at the level of changes in the phosphorylation state of tyrosyl residues in intracellular proteins.  
         [0027]    Transformed cells are transfected with the PTPH 1  DNA using an appropriate expression vector. The ability of PTPH 1  DNA or its functional equivalent to have the desired effect is assessed by determining whether the transformed phenotype is maintained or reversed. Reversal of the phenotype is indicative of the ability of the introduced DNA to counter the effects of the kinase.  
         [0028]    Thus, an agent and a method for preventing or reversing the malignant transformation of cells associated with or caused by the activity of a protein tyrosine kinase are now available. The agent can be DNA or RNA encoding PTPH 1  or its functional equivalent. A functional equivalent of PTPH 1  has substantially the same sequence as that shown in FIG. 1, and can catalyze dephosphorylation of tyrosyl residues phosphorylated through the action of a protein tyrosine kinase. As used herein, the term PTPH 1  includes the protein whose amino acid sequence is represented in FIG. 1 and its functional equivalents, as defined above. Modified or mutated PTPH 1 -encoding DNA (i.e., DNA which differs from the PTPH 1  DNA of FIG. 1 by at least one addition, deletion or substitution) can also be used.  
         [0029]    In the method of the present invention, DNA encoding PTPH 1  or its functional equivalent is introduced into an individual in whom its effects are desired. It is introduced in a recombinant vehicle, such as, but not limited to, retrovirus or vaccinia virus. All or a portion of the PTPH 1  DNA or RNA can be used, provided that it is sufficient to express in host cells a protein or peptide with the PTPase activity. PTPH 1 -encoding DNA can be introduced into a retroviral vector, such as those described in the literature, using known methods (See, for example, Yu et al.,  Proc. Natl. Acad. Sci USA,  83:3194-3198 (1986); Markowitz et al.,  J. Virol.,  62:1120-1124 (1988); Cepko et al.,  Cell,  37:1053-1062 (1984); Mann et al.,  Cell,  33:153-159 (1983); Cone et al.,  Proc. Natl. Acad. Sci. USA,  81:6349-6353 (1984)). Alternatively, it can be introduced into vaccinia virus, using known methods.  
         [0030]    The resulting recombinant gene transfer vehicle, which contains all or a portion of the PTPH 1  DNA (or RNA), is introduced into an individual in sufficient quantities to prevent malignant transformation associated with phosphorylation of tyrosyl residues in proteins via a protein tyrosine kinase-catalyzed reaction. The PTPH 1  DNA-containing vehicle can be introduced by a variety of routes (e.g., intraperitoneally, intramuscularly, intravascularly) and will generally be introduced in combination with a carrier (e.g., physiological saline or suitable buffer). This method is useful in the treatment or prevention of a variety of malignancies, including breast cancer and leukemia.  
         [0031]    EXEMPLIFICATION  
         [0032]    Identification of PTPH 1   
         [0033]    Oligonucleotide primers were synthesized based on the conserved sequences KCAQYWP (#1, equivalent to residues 120-126 in PTP 1 B) and HCSAGIG (#2, equivalent to residues 214-220 in PTP 1 B). The degeneracy of the first primer was 64 fold and that of the second 96 fold. Phage DNA of a HeLa cell cDNA library (Stratagene) isolated by the plate lysate method (Sambrook, J. et al.,  Molecular Cloning: A Lab Manual,  2d (1989)) was used as template. Primers were phosphorylated with T4 polynucleotide kinase (New England Biolabs) prior to PCR and were added at a final concentration of 1 μM to a mixture containing 10 mM Tris-HCl (pH8.4), 50 mM KCl, 1.5 mM MgCl 2 , 0.01% gelatin, 20 mM dNTPS, 2.5 units of Taq polymerase (Perkin-Elmer/Cetus) and 2 μg phage DNA. Thirty cycles of the PCR reaction were performed; each was carried out at 94° C. for 1 min, 50° C. for 1 min and 72° C. for 2 min. The PCR products were analyzed on a 1% agarose gel and DNA fragments of approximately 0.25 kb were excised, eluted from the gel, subcloned into the Smal site of pUC118 and sequenced. The same HeLa EDNA library was probed with the 0.25 kb PTPH 1  product of PCR obtained above. Plaques were transferred to nitrocellulose filters (Schleicher &amp; Schuell) and screened by hybridization at 65° C. in a solution containing 2× SSC (SSC=0.15 M NaCl, 0.015 M trisodium citrate), 5× Denhardt&#39;s, 0.1% SDS, 25 mM sodium phosphate, 1% sodium pyrophosphate, 10% dextran sulfate and 12.5 μg/ml denatured calf thymus DNA. The filters were washed successively at the same temperature in a series of solutions containing 0.1% SDS with 4× SSC, 2× SSC, 1× SSC and finally 0.1× SSC. Hybridizing phage were plaque purified, DNA was prepared and cDNA inserts were isolated and subcloned using standard techniques (Sambrook, J. et al.,  Molecular Cloning: A Lab Manual,  2d (1989)). DNA sequencing was carried out by the dideoxynucleotide chain-termination procedure (Sanger, F. et al.,  Proc. Natl. Acad. Sci. USA,  74:5463-5467 (1977)) using either manufacturer&#39;s primers (United States Biochemical Corporation) or synthetic oligonucleotides derived from the existing sequences. The GenBank database of sequence information was searched with the FASTA program of Pearson and Lipman to identify proteins with amino acid sequence similar to that of PTPH 1 .  
         [0034]    The nucleotide sequence of the cDNA designated PTPH 1  and the predicted amino acid sequence are represented in FIG. 1. The open box delineates the segment of homology to the N-terminal domain of band 4.1, ezrin and talin. The shaded box defines the segment homologous to the catalytic domains of the PTPases. In the intervening segment, seryl and threonyl residues located in sequences bearing features of sites of casein kinase II phosphorylation sites are underlined. The primary structure requirements for casein kinase II phosphorylation include the presence of surrounding acidic residues. An Asp or Glu three residues to the C-terminus of the phosphate acceptor site is particularly critical (Kuenzel, E. A. et al.,  J. Biol. Chem.,  262:9136-9140 (1987)). The presence of prolyl residues preceeding the phosphorylation site is also common. In addition, seryl residues 372 and 381 display some features of potential sites of phosphorylation by p34 cdc2  (Moreno, S. and P. Nurse,  Cell  61:549-551 (1990)).  
         [0035]    Northern analysis of PTPH 1  mRNA was also carried out. Total RNA was extracted from HeLa cells and Poly(A)+ RNA was selected by oligo(dT)-cellulose chromatography as described by Sambrook, J. et al. (Sambrook, J. et al.,  Molecular Cloning: A Lab Manual,  2d (1989)). After electrophoresis on a formaldehyde-agarose (1%) gel, the RNA was transferred to a Gene Screen Plus membrane and hybridized with the 4 kb insert of PTPH 1 . The hybridization and washing conditions were the same as described above, except that SDS was included at 1%. Results are shown in FIG. 2. Numbers on the right indicate the size of RNA markers (BRL).  
         [0036]    The relationship of the structure of PTPH 1  was assessed and compared with that of other proteins. This assessment showed that PTPH 1  comprises three segments, as represented in FIG. 3: an N-terminal segment (shaded) with homology to the membrane localization domains in band 4.1, ezrin and talin, a putative regulatory segment in the middle of the protein (open) and a C-terminal PTPase related segment (black). TM denotes the transmembrane domain in CD45. The length of each protein is shown in proportion to its molecular weight. The homology among domains is indicated by their percentage identity expressed as the number of identities out of possible matches between residues in the aligned sequences. In addition the numbers in parentheses are alignment scores, which express the similarity of segments in units of standard deviation from the average background scores of 100 randomly generated sequences. The alignments were optimised using the ALIGN programme from the National Biomedical Research Foundation, the mutation data matrix and a gap penalty of 10. Scores of &gt;5 suggest homology. Residues 3-357 in PTPH 1  were compared pairwise with 2-323 in band 4.1, 4-339 in ezrin and 120-455 in talin. Residues 669-913 in PTPH 1  were compared pairwise with 40-289 in PTP 1 B, 42-273 in TCPTPase, 491-738 in CD45 domain I and 782-1054 in CD45 domain II.  
         [0037]    EQUIVALENTS  
         [0038]    Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.  
     
       
       
         1 
         
           
             13  
           
           
             1  
             3984  
             DNA  
             Homosapiens  
             
               CDS  
               (24)...(2765)  
             
           
            1 

ctgcaggtta ttcagcgata gtt atg acc tcc cgg tta cgt gcg ttg ggt gga     53 
                          Met Thr Ser Arg Leu Arg Ala Leu Gly Gly 
                           1               5                   10 

aga att aat aat ata cgc acc tcg gag tta ccc aaa gag aaa act cga      101 
Arg Ile Asn Asn Ile Arg Thr Ser Glu Leu Pro Lys Glu Lys Thr Arg 
                 15                  20                  25 

tca gaa gtc att tgc agc atc cac ttt tta gat ggc gtg gta cag acc      149 
Ser Glu Val Ile Cys Ser Ile His Phe Leu Asp Gly Val Val Gln Thr 
             30                  35                  40 

ttt aaa gtt act aaa caa gac act ggc cag gtt ctt ctg gat atg gtg      197 
Phe Lys Val Thr Lys Gln Asp Thr Gly Gln Val Leu Leu Asp Met Val 
         45                  50                  55 

cac aac cac ctg ggt gtg act gaa aag gaa tat ttt ggt tta cag cat      245 
His Asn His Leu Gly Val Thr Glu Lys Glu Tyr Phe Gly Leu Gln His 
     60                  65                  70 

gat gac gac tcc gtg gac tct cct aga tgg ctg gaa gca agc aaa ccc      293 
Asp Asp Asp Ser Val Asp Ser Pro Arg Trp Leu Glu Ala Ser Lys Pro 
 75                  80                  85                  90 

atc agg aag cag tta aaa gga ggt ttc ccc tgt acc ctg cat ttt cga      341 
Ile Arg Lys Gln Leu Lys Gly Gly Phe Pro Cys Thr Leu His Phe Arg 
                 95                 100                 105 

gta aga ttt ttt ata cct gat ccc aac aca ctg cag caa gaa caa acc      389 
Val Arg Phe Phe Ile Pro Asp Pro Asn Thr Leu Gln Gln Glu Gln Thr 
            110                 115                 120 

agg cac ttg tat ttc tta caa ctg aag atg gat att tgc gaa gga agg      437 
Arg His Leu Tyr Phe Leu Gln Leu Lys Met Asp Ile Cys Glu Gly Arg 
        125                 130                 135 

tta acc tgc cct ctt aac tca gca gtg gtt cta gcg tcc tat gcc gta      485 
Leu Thr Cys Pro Leu Asn Ser Ala Val Val Leu Ala Ser Tyr Ala Val 
    140                 145                 150 

caa tct cat ttt gga gac tat aat tct tcc ata cat cat cca ggc tat      533 
Gln Ser His Phe Gly Asp Tyr Asn Ser Ser Ile His His Pro Gly Tyr 
155                 160                 165                 170 

ctt tcc gat agt cac ttt ata ccc gat caa aat gag gac ttt tta aca      581 
Leu Ser Asp Ser His Phe Ile Pro Asp Gln Asn Glu Asp Phe Leu Thr 
                175                 180                 185 

aaa gtc gaa tct ctg cat gag cag cac agt ggg cta aaa caa tca gaa      629 
Lys Val Glu Ser Leu His Glu Gln His Ser Gly Leu Lys Gln Ser Glu 
            190                 195                 200 

gca gaa tcc tgc tat atc aac ata gcg cgg acc ctc gac ttc tat gga      677 
Ala Glu Ser Cys Tyr Ile Asn Ile Ala Arg Thr Leu Asp Phe Tyr Gly 
        205                 210                 215 

gta gaa ctg cac agt ggt agg gat ctg cac aat tta gac cta atg att      725 
Val Glu Leu His Ser Gly Arg Asp Leu His Asn Leu Asp Leu Met Ile 
    220                 225                 230 

gga att gct tcc gcg ggt gtt gct gtg tac cga aaa tac att tgc aca      773 
Gly Ile Ala Ser Ala Gly Val Ala Val Tyr Arg Lys Tyr Ile Cys Thr 
235                 240                 245                 250 

agt ttc tat cct tgg gtg aac att ctc aaa att tct ttc aaa agg aaa      821 
Ser Phe Tyr Pro Trp Val Asn Ile Leu Lys Ile Ser Phe Lys Arg Lys 
                255                 260                 265 

aag ttc ttc ata cat cag cga cag aaa cag gct gaa tcc agg gaa cat      869 
Lys Phe Phe Ile His Gln Arg Gln Lys Gln Ala Glu Ser Arg Glu His 
            270                 275                 280 

att gtg gcc ttc aac atg ctg aat tac cga tct tgc aaa aac ttg tgg      917 
Ile Val Ala Phe Asn Met Leu Asn Tyr Arg Ser Cys Lys Asn Leu Trp 
        285                 290                 295 

aaa tcc tgt gtt gag cac cat acg ttc ttt cag gca aag aag cta cta      965 
Lys Ser Cys Val Glu His His Thr Phe Phe Gln Ala Lys Lys Leu Leu 
    300                 305                 310 

cct cag gaa aag aat gtt ctg tct cag tac tgg act atg ggc tct cgg     1013 
Pro Gln Glu Lys Asn Val Leu Ser Gln Tyr Trp Thr Met Gly Ser Arg 
315                 320                 325                 330 

aac acc aaa aag tcg gta aat aac caa tat tgc aaa aag gtg att ggc     1061 
Asn Thr Lys Lys Ser Val Asn Asn Gln Tyr Cys Lys Lys Val Ile Gly 
                335                 340                 345 

ggg atg gtg tgg aac cca gcc atg cgg aga tcc tta tca gtg gag cac     1109 
Gly Met Val Trp Asn Pro Ala Met Arg Arg Ser Leu Ser Val Glu His 
            350                 355                 360 

tta gaa acc aag agt ctg cct tct cgt tcc cct ccc att act ccc aac     1157 
Leu Glu Thr Lys Ser Leu Pro Ser Arg Ser Pro Pro Ile Thr Pro Asn 
        365                 370                 375 

tgg cga agt cct cgg ctc cgg cac gaa atc cga aag cca cgc cac tct     1205 
Trp Arg Ser Pro Arg Leu Arg His Glu Ile Arg Lys Pro Arg His Ser 
    380                 385                 390 

tct gca gat aac ctt gca aat gaa atg acc tac atc acg gaa acg gaa     1253 
Ser Ala Asp Asn Leu Ala Asn Glu Met Thr Tyr Ile Thr Glu Thr Glu 
395                 400                 405                 410 

gat gta ttt tac acg tac aag ggc tct ctg gcc cct caa gac agc gat     1301 
Asp Val Phe Tyr Thr Tyr Lys Gly Ser Leu Ala Pro Gln Asp Ser Asp 
                415                 420                 425 

tct gaa gtt tct cag aac cga agc ccg cac caa gag agt tta tcc gag     1349 
Ser Glu Val Ser Gln Asn Arg Ser Pro His Gln Glu Ser Leu Ser Glu 
            430                 435                 440 

aac aat ccg gca caa agc tac ctg acc cag aag tca tcc agt tct gtg     1397 
Asn Asn Pro Ala Gln Ser Tyr Leu Thr Gln Lys Ser Ser Ser Ser Val 
        445                 450                 455 

tct cca tct tca aat gct cca ggc tcc tgc tca cct gac ggc gtt gat     1445 
Ser Pro Ser Ser Asn Ala Pro Gly Ser Cys Ser Pro Asp Gly Val Asp 
    460                 465                 470 

cag cag ctc tta gat gac ttc cac agg gtg acc aaa ggg ggc tcc acc     1493 
Gln Gln Leu Leu Asp Asp Phe His Arg Val Thr Lys Gly Gly Ser Thr 
475                 480                 485                 490 

gag gac gcc agc cag tac tac tgt gac aag aat gat aat ggt gac agc     1541 
Glu Asp Ala Ser Gln Tyr Tyr Cys Asp Lys Asn Asp Asn Gly Asp Ser 
                495                 500                 505 

tac tta gtc ttg atc cgt atc aca cca gat gaa gat gga aaa ttt gga     1589 
Tyr Leu Val Leu Ile Arg Ile Thr Pro Asp Glu Asp Gly Lys Phe Gly 
            510                 515                 520 

ttt aat ctt aag gga gga gtg gat caa aag atg cct ctt gtg gta tca     1637 
Phe Asn Leu Lys Gly Gly Val Asp Gln Lys Met Pro Leu Val Val Ser 
        525                 530                 535 

agg ata aac cca gag tca cct gcg gac acc tgc att cct aag ctg aac     1685 
Arg Ile Asn Pro Glu Ser Pro Ala Asp Thr Cys Ile Pro Lys Leu Asn 
    540                 545                 550 

gaa ggg gat caa atc gtg tta atc aat ggc cgg gac atc tca gaa cac     1733 
Glu Gly Asp Gln Ile Val Leu Ile Asn Gly Arg Asp Ile Ser Glu His 
555                 560                 565                 570 

acg cat gac caa gtg gtg atg ttc atc aaa gcc agc cgg gag tcc cac     1781 
Thr His Asp Gln Val Val Met Phe Ile Lys Ala Ser Arg Glu Ser His 
                575                 580                 585 

tca cgg gag ctg gcc ctg gtg atc agg agg aga gct gtc cgc tca ttt     1829 
Ser Arg Glu Leu Ala Leu Val Ile Arg Arg Arg Ala Val Arg Ser Phe 
            590                 595                 600 

gct gac ttc aag tct gaa gat gaa ctg aac cag ctt ttc ccc gaa gcc     1877 
Ala Asp Phe Lys Ser Glu Asp Glu Leu Asn Gln Leu Phe Pro Glu Ala 
        605                 610                 615 

att ttc ccc atg tgt ccg gag ggt ggg gac act ttg gag gga tcc atg     1925 
Ile Phe Pro Met Cys Pro Glu Gly Gly Asp Thr Leu Glu Gly Ser Met 
    620                 625                 630 

gca cag cta aag aag ggc ctc gaa agc ggg acg gtg ctg atc cag ttt     1973 
Ala Gln Leu Lys Lys Gly Leu Glu Ser Gly Thr Val Leu Ile Gln Phe 
635                 640                 645                 650 

gag caa ctc tac aga aaa aag cca ggt ttg gcc atc acg ttt gca aag     2021 
Glu Gln Leu Tyr Arg Lys Lys Pro Gly Leu Ala Ile Thr Phe Ala Lys 
                655                 660                 665 

ctg cct caa aat ttg gac aaa aac cga tat aaa gat gtg ctg cct tat     2069 
Leu Pro Gln Asn Leu Asp Lys Asn Arg Tyr Lys Asp Val Leu Pro Tyr 
            670                 675                 680 

gac acc acc cgg gta tta ttg cag gga aat gaa gat tat att aat gca     2117 
Asp Thr Thr Arg Val Leu Leu Gln Gly Asn Glu Asp Tyr Ile Asn Ala 
        685                 690                 695 

agt tac gtg aac atg gaa att cct gct gct aac ctt gtg aac aag tac     2165 
Ser Tyr Val Asn Met Glu Ile Pro Ala Ala Asn Leu Val Asn Lys Tyr 
    700                 705                 710 

atc gcc act cag ggg ccc ctg ccg cat acc tgt gca cag ttt tgg cag     2213 
Ile Ala Thr Gln Gly Pro Leu Pro His Thr Cys Ala Gln Phe Trp Gln 
715                 720                 725                 730 

gtt gtc tgg gat cag aag ttg tca ctc att gtc atg ttg acg act ctc     2261 
Val Val Trp Asp Gln Lys Leu Ser Leu Ile Val Met Leu Thr Thr Leu 
                735                 740                 745 

aca gaa cga ggg cgg acc aaa tgt cac cag tac tgg cca gat ccc ccc     2309 
Thr Glu Arg Gly Arg Thr Lys Cys His Gln Tyr Trp Pro Asp Pro Pro 
            750                 755                 760 

gac gtc atg aac cac ggc ggc ttt cac atc cag tgt cag tca gag gac     2357 
Asp Val Met Asn His Gly Gly Phe His Ile Gln Cys Gln Ser Glu Asp 
        765                 770                 775 

tgc acc atc gcc tat gtg tcc cga gaa atg ctg gtc aca aac acc cag     2405 
Cys Thr Ile Ala Tyr Val Ser Arg Glu Met Leu Val Thr Asn Thr Gln 
    780                 785                 790 

acc ggg gaa gaa cac aca gtg aca cat ctc cag tac gtc gca tgg cct     2453 
Thr Gly Glu Glu His Thr Val Thr His Leu Gln Tyr Val Ala Trp Pro 
795                 800                 805                 810 

gac cac ggt ata ccc gat gac tcc tcc gac ttt ctg gaa ttt gta aac     2501 
Asp His Gly Ile Pro Asp Asp Ser Ser Asp Phe Leu Glu Phe Val Asn 
                815                 820                 825 

tat gtg agg tct ctg aga gtg gac agc gag cct gtc cta gtt cac tgc     2549 
Tyr Val Arg Ser Leu Arg Val Asp Ser Glu Pro Val Leu Val His Cys 
            830                 835                 840 

agt gct gga ata ggt cga acc ggt gtg ttg gtc act atg gaa aca gcc     2597 
Ser Ala Gly Ile Gly Arg Thr Gly Val Leu Val Thr Met Glu Thr Ala 
        845                 850                 855 

atg tgc cta act gag agg aac ctg ccc att tac cca ctg gat att gtc     2645 
Met Cys Leu Thr Glu Arg Asn Leu Pro Ile Tyr Pro Leu Asp Ile Val 
    860                 865                 870 

cga aaa atg cga gac cag cgc gcc atg atg gtg cag aca tca agc cag     2693 
Arg Lys Met Arg Asp Gln Arg Ala Met Met Val Gln Thr Ser Ser Gln 
875                 880                 885                 890 

tac aag ttt gtg tgt gaa gcg att ctt cgt gtg tat gaa gaa ggt tta     2741 
Tyr Lys Phe Val Cys Glu Ala Ile Leu Arg Val Tyr Glu Glu Gly Leu 
                895                 900                 905 

gtc caa atg ctg gat cct agt taa gacaactgtg aaaaagttca ttcctctttc    2795 
Val Gln Met Leu Asp Pro Ser  * 
            910 

ccaagggcat cctccttgaa agaggaggac agacctctct ggaagcagca agaggaacca   2855 

gtagctgtgg gaaaggaatg ggcacctctg aacccaggca ctttaaactt ctatagaaaa   2915 

gatatcgtgt acataggaac tggtgtagat aagcatgcaa ttatggcatc atttaggcct   2975 

gtatttctat ggaaagatac aaaaaggatc tcagtttggg gcctgtccta atgccttctt   3035 

ccctaacatc accacacaca cccctgtcgg catcctggag caattgagac cggacaccca   3095 

cagagctgtt gtcctcccag caacaagatg gtgtggttat cttgggtcat ttggatgttt   3155 

tgtttgtttc tgtgtgtcag actgtaaggg ctgagctttc tgtgcttcta ggtggagctg   3215 

gaacaattca gattcacccg ccctgatgct aaggaaaccc tgacgtatgt actagatggc   3275 

agggcactgg gggtcaggct gaaggctgag caacacctct ctgccctccc tccctttgtc   3335 

ccatctccca gcgacttcca atattcatgt ttctgagaat tgtgtccctc ttcagttccc   3395 

tcttggtgcc taacctggat tagtaatgtg cattcaggtg aattttcagc tgaggctctg   3455 

agaactggta ctctcagtgt gttctggtca tcttgtggct tagttgtaga agcaggtgtg   3515 

tctcttgcct ctgcttgcct cctactgcac actcagcacc caggactgga atcaccgact   3575 

actgaatctc ctacatgtat tgctgctact tcaagctcct ccacttgaaa ccttatgatt   3635 

ttccaagggg agatgggaca gtgtcatcta aatattccga atgtttggcc ttctgagaaa   3695 

agagcttcta gtaattgaac catgggtttc ccagcttctg gagggttggc cgtgggctgt   3755 

gtacatgtgt gtgcccaggg gtgagtgttt ctcaggattc ctaacgattc aaattaccgt   3815 

tgagtatata taaagaatcg agtcggaaga acaaatgtgt gcattcaccc ccagtcacaa   3875 

tggtctccat tgcatttcaa aggagaggat cagactatct gaatataaac acaatctgat   3935 

gttaatttat tctaagaaca ccatctgtat tcattttgat tgtcctaaa               3984 

 
           
             2  
             913  
             PRT  
             Homosapiens  
           
            2 

Met Thr Ser Arg Leu Arg Ala Leu Gly Gly Arg Ile Asn Asn Ile Arg 
 1               5                  10                  15 

Thr Ser Glu Leu Pro Lys Glu Lys Thr Arg Ser Glu Val Ile Cys Ser 
            20                  25                  30 

Ile His Phe Leu Asp Gly Val Val Gln Thr Phe Lys Val Thr Lys Gln 
        35                  40                  45 

Asp Thr Gly Gln Val Leu Leu Asp Met Val His Asn His Leu Gly Val 
    50                  55                  60 

Thr Glu Lys Glu Tyr Phe Gly Leu Gln His Asp Asp Asp Ser Val Asp 
65                  70                  75                  80 

Ser Pro Arg Trp Leu Glu Ala Ser Lys Pro Ile Arg Lys Gln Leu Lys 
                85                  90                  95 

Gly Gly Phe Pro Cys Thr Leu His Phe Arg Val Arg Phe Phe Ile Pro 
            100                 105                 110 

Asp Pro Asn Thr Leu Gln Gln Glu Gln Thr Arg His Leu Tyr Phe Leu 
        115                 120                 125 

Gln Leu Lys Met Asp Ile Cys Glu Gly Arg Leu Thr Cys Pro Leu Asn 
    130                 135                 140 

Ser Ala Val Val Leu Ala Ser Tyr Ala Val Gln Ser His Phe Gly Asp 
145                 150                 155                 160 

Tyr Asn Ser Ser Ile His His Pro Gly Tyr Leu Ser Asp Ser His Phe 
                165                 170                 175 

Ile Pro Asp Gln Asn Glu Asp Phe Leu Thr Lys Val Glu Ser Leu His 
            180                 185                 190 

Glu Gln His Ser Gly Leu Lys Gln Ser Glu Ala Glu Ser Cys Tyr Ile 
        195                 200                 205 

Asn Ile Ala Arg Thr Leu Asp Phe Tyr Gly Val Glu Leu His Ser Gly 
    210                 215                 220 

Arg Asp Leu His Asn Leu Asp Leu Met Ile Gly Ile Ala Ser Ala Gly 
225                 230                 235                 240 

Val Ala Val Tyr Arg Lys Tyr Ile Cys Thr Ser Phe Tyr Pro Trp Val 
                245                 250                 255 

Asn Ile Leu Lys Ile Ser Phe Lys Arg Lys Lys Phe Phe Ile His Gln 
            260                 265                 270 

Arg Gln Lys Gln Ala Glu Ser Arg Glu His Ile Val Ala Phe Asn Met 
        275                 280                 285 

Leu Asn Tyr Arg Ser Cys Lys Asn Leu Trp Lys Ser Cys Val Glu His 
    290                 295                 300 

His Thr Phe Phe Gln Ala Lys Lys Leu Leu Pro Gln Glu Lys Asn Val 
305                 310                 315                 320 

Leu Ser Gln Tyr Trp Thr Met Gly Ser Arg Asn Thr Lys Lys Ser Val 
                325                 330                 335 

Asn Asn Gln Tyr Cys Lys Lys Val Ile Gly Gly Met Val Trp Asn Pro 
            340                 345                 350 

Ala Met Arg Arg Ser Leu Ser Val Glu His Leu Glu Thr Lys Ser Leu 
        355                 360                 365 

Pro Ser Arg Ser Pro Pro Ile Thr Pro Asn Trp Arg Ser Pro Arg Leu 
    370                 375                 380 

Arg His Glu Ile Arg Lys Pro Arg His Ser Ser Ala Asp Asn Leu Ala 
385                 390                 395                 400 

Asn Glu Met Thr Tyr Ile Thr Glu Thr Glu Asp Val Phe Tyr Thr Tyr 
                405                 410                 415 

Lys Gly Ser Leu Ala Pro Gln Asp Ser Asp Ser Glu Val Ser Gln Asn 
            420                 425                 430 

Arg Ser Pro His Gln Glu Ser Leu Ser Glu Asn Asn Pro Ala Gln Ser 
        435                 440                 445 

Tyr Leu Thr Gln Lys Ser Ser Ser Ser Val Ser Pro Ser Ser Asn Ala 
    450                 455                 460 

Pro Gly Ser Cys Ser Pro Asp Gly Val Asp Gln Gln Leu Leu Asp Asp 
465                 470                 475                 480 

Phe His Arg Val Thr Lys Gly Gly Ser Thr Glu Asp Ala Ser Gln Tyr 
                485                 490                 495 

Tyr Cys Asp Lys Asn Asp Asn Gly Asp Ser Tyr Leu Val Leu Ile Arg 
            500                 505                 510 

Ile Thr Pro Asp Glu Asp Gly Lys Phe Gly Phe Asn Leu Lys Gly Gly 
        515                 520                 525 

Val Asp Gln Lys Met Pro Leu Val Val Ser Arg Ile Asn Pro Glu Ser 
    530                 535                 540 

Pro Ala Asp Thr Cys Ile Pro Lys Leu Asn Glu Gly Asp Gln Ile Val 
545                 550                 555                 560 

Leu Ile Asn Gly Arg Asp Ile Ser Glu His Thr His Asp Gln Val Val 
                565                 570                 575 

Met Phe Ile Lys Ala Ser Arg Glu Ser His Ser Arg Glu Leu Ala Leu 
            580                 585                 590 

Val Ile Arg Arg Arg Ala Val Arg Ser Phe Ala Asp Phe Lys Ser Glu 
        595                 600                 605 

Asp Glu Leu Asn Gln Leu Phe Pro Glu Ala Ile Phe Pro Met Cys Pro 
    610                 615                 620 

Glu Gly Gly Asp Thr Leu Glu Gly Ser Met Ala Gln Leu Lys Lys Gly 
625                 630                 635                 640 

Leu Glu Ser Gly Thr Val Leu Ile Gln Phe Glu Gln Leu Tyr Arg Lys 
                645                 650                 655 

Lys Pro Gly Leu Ala Ile Thr Phe Ala Lys Leu Pro Gln Asn Leu Asp 
            660                 665                 670 

Lys Asn Arg Tyr Lys Asp Val Leu Pro Tyr Asp Thr Thr Arg Val Leu 
        675                 680                 685 

Leu Gln Gly Asn Glu Asp Tyr Ile Asn Ala Ser Tyr Val Asn Met Glu 
    690                 695                 700 

Ile Pro Ala Ala Asn Leu Val Asn Lys Tyr Ile Ala Thr Gln Gly Pro 
705                 710                 715                 720 

Leu Pro His Thr Cys Ala Gln Phe Trp Gln Val Val Trp Asp Gln Lys 
                725                 730                 735 

Leu Ser Leu Ile Val Met Leu Thr Thr Leu Thr Glu Arg Gly Arg Thr 
            740                 745                 750 

Lys Cys His Gln Tyr Trp Pro Asp Pro Pro Asp Val Met Asn His Gly 
        755                 760                 765 

Gly Phe His Ile Gln Cys Gln Ser Glu Asp Cys Thr Ile Ala Tyr Val 
    770                 775                 780 

Ser Arg Glu Met Leu Val Thr Asn Thr Gln Thr Gly Glu Glu His Thr 
785                 790                 795                 800 

Val Thr His Leu Gln Tyr Val Ala Trp Pro Asp His Gly Ile Pro Asp 
                805                 810                 815 

Asp Ser Ser Asp Phe Leu Glu Phe Val Asn Tyr Val Arg Ser Leu Arg 
            820                 825                 830 

Val Asp Ser Glu Pro Val Leu Val His Cys Ser Ala Gly Ile Gly Arg 
        835                 840                 845 

Thr Gly Val Leu Val Thr Met Glu Thr Ala Met Cys Leu Thr Glu Arg 
    850                 855                 860 

Asn Leu Pro Ile Tyr Pro Leu Asp Ile Val Arg Lys Met Arg Asp Gln 
865                 870                 875                 880 

Arg Ala Met Met Val Gln Thr Ser Ser Gln Tyr Lys Phe Val Cys Glu 
                885                 890                 895 

Ala Ile Leu Arg Val Tyr Glu Glu Gly Leu Val Gln Met Leu Asp Pro 
            900                 905                 910 

Ser 

 
           
             3  
             328  
             PRT  
             Homosapiens  
           
            3 

Ile Cys Ser Ile His Phe Leu Asp Gly Val Val Gln Thr Phe Lys Val 
 1               5                  10                  15 

Thr Lys Gln Asp Thr Gly Gln Val Leu Leu Asp Met Val His Asn His 
            20                  25                  30 

Leu Gly Val Thr Glu Lys Glu Tyr Phe Gly Leu Gln His Asp Asp Asp 
        35                  40                  45 

Ser Val Asp Ser Pro Arg Trp Leu Glu Ala Ser Lys Pro Ile Arg Lys 
    50                  55                  60 

Gln Leu Lys Gly Gly Phe Pro Cys Thr Leu His Phe Arg Val Arg Phe 
65                  70                  75                  80 

Phe Ile Pro Asp Pro Asn Thr Leu Gln Gln Glu Gln Thr Arg His Leu 
                85                  90                  95 

Tyr Phe Leu Gln Leu Lys Met Asp Ile Cys Glu Gly Arg Leu Thr Cys 
            100                 105                 110 

Pro Leu Asn Ser Ala Val Val Leu Ala Ser Tyr Ala Val Gln Ser His 
        115                 120                 125 

Phe Gly Asp Tyr Asn Ser Ser Ile His His Pro Gly Tyr Leu Ser Asp 
    130                 135                 140 

Ser His Phe Ile Pro Asp Gln Asn Glu Asp Phe Leu Thr Lys Val Glu 
145                 150                 155                 160 

Ser Leu His Glu Gln His Ser Gly Leu Lys Gln Ser Glu Ala Glu Ser 
                165                 170                 175 

Cys Tyr Ile Asn Ile Ala Arg Thr Leu Asp Phe Tyr Gly Val Glu Leu 
            180                 185                 190 

His Ser Gly Arg Asp Leu His Asn Leu Asp Leu Met Ile Gly Ile Ala 
        195                 200                 205 

Ser Ala Gly Val Ala Val Tyr Arg Lys Tyr Ile Cys Thr Ser Phe Tyr 
    210                 215                 220 

Pro Trp Val Asn Ile Leu Lys Ile Ser Phe Lys Arg Lys Lys Phe Phe 
225                 230                 235                 240 

Ile His Gln Arg Gln Lys Gln Ala Glu Ser Arg Glu His Ile Val Ala 
                245                 250                 255 

Phe Asn Met Leu Asn Tyr Arg Ser Cys Lys Asn Leu Trp Lys Ser Cys 
            260                 265                 270 

Val Glu His His Thr Phe Phe Gln Ala Lys Lys Leu Leu Pro Gln Glu 
        275                 280                 285 

Lys Asn Val Leu Ser Gln Tyr Trp Thr Met Gly Ser Arg Asn Thr Lys 
    290                 295                 300 

Lys Ser Val Asn Asn Gln Tyr Cys Lys Lys Val Ile Gly Gly Met Val 
305                 310                 315                 320 

Trp Asn Pro Ala Met Arg Arg Ser 
                325 

 
           
             4  
             322  
             PRT  
             Homosapiens  
           
            4 

His Cys Lys Val Ser Leu Leu Asp Asp Thr Val Tyr Glu Cys Val Val 
 1               5                  10                  15 

Glu Lys His Ala Lys Gly Gln Asp Leu Leu Lys Arg Val Cys Glu His 
            20                  25                  30 

Leu Asn Leu Leu Glu Glu Asp Tyr Phe Gly Leu Ala Ile Trp Asp Asn 
        35                  40                  45 

Ala Thr Ser Lys Thr Trp Leu Asp Ser Ala Lys Glu Ile Lys Lys Gln 
    50                  55                  60 

Val Arg Gly Val Pro Trp Asn Phe Thr Phe Asn Val Lys Phe Tyr Pro 
65                  70                  75                  80 

Pro Asp Pro Ala Gln Leu Thr Glu Asp Ile Thr Arg Tyr Tyr Leu Cys 
                85                  90                  95 

Leu Gln Leu Arg Gln Asp Ile Val Ala Gly Arg Leu Pro Cys Ser Phe 
            100                 105                 110 

Ala Thr Leu Ala Leu Leu Gly Ser Tyr Thr Ile Gln Ser Glu Leu Gly 
        115                 120                 125 

Asp Tyr Asp Pro Glu Leu His Gly Val Asp Tyr Val Ser Asp Phe Lys 
    130                 135                 140 

Leu Ala Pro Asn Gln Thr Lys Glu Leu Glu Glu Lys Val Met Glu Leu 
145                 150                 155                 160 

His Lys Ser Tyr Arg Ser Met Thr Pro Ala Gln Ala Asp Leu Glu Phe 
                165                 170                 175 

Leu Glu Asn Ala Lys Lys Leu Ser Met Tyr Gly Val Asp Leu His Lys 
            180                 185                 190 

Ala Lys Asp Leu Glu Gly Val Asp Ile Ile Leu Gly Val Cys Ser Ser 
        195                 200                 205 

Gly Leu Leu Val Tyr Lys Asp Lys Leu Arg Ile Asn Arg Phe Pro Trp 
    210                 215                 220 

Pro Lys Val Leu Lys Ile Ser Tyr Lys Arg Ser Ser Phe Phe Ile Lys 
225                 230                 235                 240 

Ile Arg Pro Gly Glu Gln Glu Gln Tyr Glu Ser Thr Ile Gly Phe Lys 
                245                 250                 255 

Leu Pro Ser Tyr Arg Ala Ala Lys Lys Leu Trp Lys Val Cys Val Glu 
            260                 265                 270 

His His Thr Thr Phe Arg Leu Thr Ser Thr Asp Thr Ile Pro Lys Ser 
        275                 280                 285 

Lys Phe Leu Ala Leu Gly Ser Lys Phe Arg Tyr Ser Gly Arg Thr Gln 
    290                 295                 300 

Ala Gln Thr Arg Gln Ala Ser Ala Leu Ile Asp Arg Pro Ala Pro His 
305                 310                 315                 320 

Phe Glu 

 
           
             5  
             336  
             PRT  
             Homosapiens  
           
            5 

Ile Asn Val Arg Val Thr Thr Met Asp Ala Glu Leu Glu Phe Ala Ile 
 1               5                  10                  15 

Gln Pro Asn Thr Thr Gly Lys Gln Leu Phe Asp Gln Val Val Lys Thr 
            20                  25                  30 

Ile Gly Leu Arg Glu Val Trp Tyr Phe Gly Leu His Tyr Val Asp Asn 
        35                  40                  45 

Lys Gly Phe Pro Thr Trp Leu Lys Leu Asp Lys Lys Val Ser Ala Gln 
    50                  55                  60 

Glu Val Arg Lys Glu Asn Pro Leu Gln Phe Lys Phe Arg Ala Lys Phe 
65                  70                  75                  80 

Tyr Pro Glu Asp Val Ala Glu Glu Leu Ile Gln Asp Ile Thr Gln Lys 
                85                  90                  95 

Leu Phe Phe Leu Gln Val Lys Glu Gly Ile Leu Ser Asp Glu Ile Tyr 
            100                 105                 110 

Cys Pro Pro Glu Thr Ala Val Leu Leu Gly Ser Tyr Ala Val Gln Ala 
        115                 120                 125 

Lys Phe Gly Asp Tyr Asn Lys Glu Val His Lys Ser Gly Tyr Leu Ser 
    130                 135                 140 

Ser Glu Arg Leu Ile Pro Gln Arg Val Met Asp Gln His Lys Leu Thr 
145                 150                 155                 160 

Arg Asp Gln Trp Glu Asp Arg Ile Gln Val Trp His Ala Glu His Arg 
                165                 170                 175 

Gly Met Leu Lys Asp Asn Ala Met Leu Glu Tyr Leu Lys Ile Ala Gln 
            180                 185                 190 

Asp Leu Glu Met Tyr Gly Ile Asn Tyr Phe Glu Ile Lys Asn Lys Lys 
        195                 200                 205 

Gly Thr Asp Leu Trp Leu Gly Val Asp Ala Leu Gly Leu Asn Ile Tyr 
    210                 215                 220 

Glu Lys Asp Asp Lys Leu Thr Pro Lys Ile Gly Phe Pro Trp Ser Glu 
225                 230                 235                 240 

Ile Arg Asn Ile Ser Phe Asn Asp Lys Lys Phe Val Ile Lys Pro Ile 
                245                 250                 255 

Asp Lys Lys Ala Pro Asp Phe Val Phe Tyr Ala Pro Arg Leu Arg Ile 
            260                 265                 270 

Asn Lys Arg Ile Leu Gln Leu Cys Met Gly Asn His Glu Leu Tyr Met 
        275                 280                 285 

Arg Arg Arg Lys Pro Asp Thr Ile Glu Val Gln Gln Met Lys Ala Gln 
    290                 295                 300 

Ala Arg Glu Glu Lys His Gln Lys Gln Leu Glu Arg Gln Gln Leu Glu 
305                 310                 315                 320 

Thr Glu Lys Lys Arg Arg Glu Thr Val Glu Arg Glu Lys Glu Gln Met 
                325                 330                 335 

 
           
             6  
             336  
             PRT  
             Mus musculus  
           
            6 

Gly Ile Thr Asn His Asp Glu Tyr Ser Leu Val Arg Glu Leu Met Glu 
 1               5                  10                  15 

Glu Lys Lys Asp Glu Gly Thr Gly Thr Leu Arg Lys Asp Lys Thr Leu 
            20                  25                  30 

Leu Arg Asp Glu Lys Lys Met Glu Lys Leu Lys Gln Lys Leu His Thr 
        35                  40                  45 

Asp Asp Glu Leu Asn Trp Leu Asp His Gly Arg Thr Leu Arg Glu Gln 
    50                  55                  60 

Gly Val Glu Glu His Glu Thr Leu Leu Leu Arg Arg Lys Phe Phe Tyr 
65                  70                  75                  80 

Ser Asp Gln Asn Val Asp Ser Arg Asp Pro Val Gln Leu Asn Leu Leu 
                85                  90                  95 

Tyr Val Gln Ala Arg Asp Asp Ile Leu Asn Gly Ser His Pro Val Ser 
            100                 105                 110 

Pro Asp Lys Ala Cys Glu Phe Ala Gly Phe Gln Cys Gln Ile Gln Phe 
        115                 120                 125 

Gly Pro His Asn Glu Gln Lys His Lys Ala Gly Phe Leu Asp Leu Lys 
    130                 135                 140 

Asp Phe Leu Pro Lys Glu Tyr Val Lys Gln Lys Gly Glu Arg Lys Ile 
145                 150                 155                 160 

Phe Gln Ala His Lys Asn Cys Gly Gln Met Ser Glu Ile Glu Ala Lys 
                165                 170                 175 

Val Arg Tyr Val Lys Leu Ala Arg Ser Leu Lys Thr Tyr Gly Val Ser 
            180                 185                 190 

Phe Phe Leu Val Lys Glu Lys Met Lys Gly Lys Asn Lys Leu Val Pro 
        195                 200                 205 

Arg Leu Leu Gly Ile Thr Lys Glu Cys Val Met Arg Val Asp Glu Lys 
    210                 215                 220 

Thr Lys Glu Val Ile Gln Glu Trp Ser Leu Thr Asn Ile Lys Arg Trp 
225                 230                 235                 240 

Ala Ala Ser Pro Lys Ser Phe Thr Leu Asp Phe Gly Asp Tyr Gln Asp 
                245                 250                 255 

Gly Tyr Tyr Ser Val Gln Thr Thr Glu Gly Glu Gln Ile Ala Gln Leu 
            260                 265                 270 

Ile Ala Gly Tyr Ile Asp Ile Ile Leu Lys Lys Lys Lys Ser Lys Asp 
        275                 280                 285 

His Phe Gly Leu Glu Gly Asp Glu Glu Ser Thr Met Leu Glu Asp Ser 
    290                 295                 300 

Val Ser Pro Lys Lys Ser Thr Val Leu Gln Gln Gln Tyr Asn Arg Val 
305                 310                 315                 320 

Gly Lys Val Glu His Gly Ser Val Ala Leu Pro Ala Ile Met Arg Ser 
                325                 330                 335 

 
           
             7  
             244  
             PRT  
             Homosapiens  
           
            7 

Asn Leu Asp Lys Asn Arg Tyr Lys Asp Val Leu Pro Tyr Asp Thr Thr 
 1               5                  10                  15 

Arg Val Leu Leu Gln Gly Asn Glu Asp Tyr Ile Asn Ala Ser Tyr Val 
            20                  25                  30 

Asn Met Glu Ile Pro Ala Ala Asn Leu Val Asn Lys Tyr Ile Ala Thr 
        35                  40                  45 

Gln Gly Pro Leu Pro His Thr Cys Ala Gln Phe Trp Gln Val Val Trp 
    50                  55                  60 

Asp Gln Lys Leu Ser Leu Ile Val Met Leu Thr Thr Leu Thr Glu Arg 
65                  70                  75                  80 

Gly Arg Thr Lys Cys His Gln Tyr Trp Pro Asp Pro Pro Asp Val Met 
                85                  90                  95 

Asn His Gly Gly Phe His Ile Gln Cys Gln Ser Glu Asp Cys Thr Ile 
            100                 105                 110 

Ala Tyr Val Ser Arg Glu Met Leu Val Thr Asn Thr Gln Thr Gly Glu 
        115                 120                 125 

Glu His Thr Val Thr His Leu Gln Tyr Val Ala Trp Pro Asp His Gly 
    130                 135                 140 

Ile Pro Asp Asp Ser Ser Asp Phe Leu Glu Phe Val Asn Tyr Val Arg 
145                 150                 155                 160 

Ser Leu Arg Val Asp Ser Glu Pro Val Leu Val His Cys Ser Ala Gly 
                165                 170                 175 

Ile Gly Arg Thr Gly Val Leu Val Thr Met Glu Thr Ala Met Cys Leu 
            180                 185                 190 

Thr Glu Arg Asn Leu Pro Ile Tyr Pro Leu Asp Ile Val Arg Lys Met 
        195                 200                 205 

Arg Asp Gln Arg Ala Met Met Val Gln Thr Ser Ser Gln Tyr Lys Phe 
    210                 215                 220 

Val Cys Glu Ala Ile Leu Arg Val Tyr Glu Glu Gly Leu Val Gln Met 
225                 230                 235                 240 

Leu Asp Pro Ser 

 
           
             8  
             250  
             PRT  
             Homosapiens  
           
            8 

Asn Lys Asn Arg Asn Arg Tyr Arg Asp Val Ser Pro Phe Asp His Ser 
 1               5                  10                  15 

Arg Ile Lys Leu His Gln Glu Asp Asn Asp Tyr Ile Asn Ala Ser Leu 
            20                  25                  30 

Ile Lys Met Glu Glu Ala Gln Arg Ser Tyr Ile Leu Thr Gln Gly Pro 
        35                  40                  45 

Leu Pro Asn Thr Cys Gly His Phe Trp Glu Met Val Trp Glu Gln Lys 
    50                  55                  60 

Ser Arg Gly Val Val Met Leu Asn Arg Val Met Glu Lys Gly Ser Leu 
65                  70                  75                  80 

Lys Cys Ala Gln Tyr Trp Pro Gln Lys Glu Glu Lys Glu Met Ile Phe 
                85                  90                  95 

Glu Asp Thr Asn Leu Lys Leu Thr Leu Ile Ser Glu Asp Ile Lys Ser 
            100                 105                 110 

Tyr Tyr Thr Val Arg Gln Leu Glu Leu Glu Asn Leu Thr Thr Gln Glu 
        115                 120                 125 

Thr Arg Glu Ile Leu His Phe His Tyr Thr Thr Trp Pro Asp Phe Gly 
    130                 135                 140 

Val Pro Glu Ser Pro Ala Ser Phe Leu Asn Phe Leu Phe Lys Val Arg 
145                 150                 155                 160 

Glu Ser Gly Ser Leu Ser Pro Glu His Gly Pro Val Val Val His Cys 
                165                 170                 175 

Ser Ala Gly Ile Gly Arg Ser Gly Thr Phe Cys Leu Ala Asp Thr Cys 
            180                 185                 190 

Leu Leu Leu Met Asp Lys Arg Lys Asp Pro Ser Ser Val Asp Ile Lys 
        195                 200                 205 

Lys Val Leu Leu Glu Met Arg Lys Phe Arg Met Gly Leu Ile Gln Thr 
    210                 215                 220 

Ala Asp Gln Leu Arg Phe Ser Tyr Leu Ala Val Ile Glu Gly Ala Lys 
225                 230                 235                 240 

Phe Ile Met Gly Asp Ser Ser Val Gln Asp 
                245                 250 

 
           
             9  
             246  
             PRT  
             Homosapiens  
           
            9 

Asn Arg Asn Arg Asn Arg Tyr Arg Asp Val Ser Pro Tyr Asp His Ser 
 1               5                  10                  15 

Arg Val Lys Leu Gln Asn Ala Glu Asn Asp Tyr Ile Asn Ala Ser Leu 
            20                  25                  30 

Val Asp Ile Glu Glu Ala Gln Arg Ser Tyr Ile Leu Thr Gln Gly Pro 
        35                  40                  45 

Leu Pro Asn Thr Cys Cys His Phe Trp Leu Met Val Trp Gln Gln Lys 
    50                  55                  60 

Thr Lys Ala Val Val Met Leu Asn Arg Ile Val Glu Lys Glu Ser Val 
65                  70                  75                  80 

Lys Cys Ala Gln Tyr Trp Pro Thr Asp Asp Gln Glu Met Leu Phe Lys 
                85                  90                  95 

Glu Thr Gly Phe Ser Val Lys Leu Leu Ser Glu Asp Val Lys Ser Tyr 
            100                 105                 110 

Tyr Thr Val His Leu Leu Gln Leu Glu Asn Ile Asn Ser Gly Glu Thr 
        115                 120                 125 

Arg Thr Ile Ser His Phe His Tyr Thr Thr Trp Pro Asp Phe Gly Val 
    130                 135                 140 

Pro Glu Ser Pro Ala Ser Phe Leu Asn Phe Leu Phe Lys Val Arg Glu 
145                 150                 155                 160 

Ser Gly Ser Leu Asn Pro Asp His Gly Pro Ala Val Ile His Cys Ser 
                165                 170                 175 

Ala Gly Ile Gly Arg Ser Gly Thr Phe Ser Leu Val Asp Thr Cys Leu 
            180                 185                 190 

Val Leu Met Glu Lys Gly Asp Asp Ile Asn Ile Lys Gln Val Leu Leu 
        195                 200                 205 

Asn Met Arg Lys Tyr Arg Met Gly Leu Ile Gln Thr Pro Asp Gln Leu 
    210                 215                 220 

Arg Phe Ser Tyr Met Ala Ile Ile Glu Gly Ala Lys Cys Ile Lys Gly 
225                 230                 235                 240 

Asp Ser Ser Ile Gln Lys 
                245 

 
           
             10  
             248  
             PRT  
             Homosapiens  
           
            10 

Asn Gln Asn Lys Asn Arg Tyr Val Asp Ile Leu Pro Tyr Asp Tyr Asn 
 1               5                  10                  15 

Arg Val Glu Leu Ser Glu Ile Asn Gly Asp Ala Gly Ser Asn Tyr Ile 
            20                  25                  30 

Asn Ala Ser Tyr Ile Asp Gly Phe Lys Glu Pro Arg Lys Tyr Ile Ala 
        35                  40                  45 

Ala Gln Gly Pro Arg Asp Glu Thr Val Asp Asp Phe Trp Arg Met Ile 
    50                  55                  60 

Trp Glu Gln Lys Ala Thr Val Ile Val Met Val Thr Arg Cys Glu Glu 
65                  70                  75                  80 

Gly Asn Arg Asn Lys Cys Ala Glu Tyr Trp Pro Ser Met Glu Glu Gly 
                85                  90                  95 

Thr Arg Ala Phe Gly Asp Val Val Val Lys Ile Asn Gln His Lys Arg 
            100                 105                 110 

Cys Pro Asp Tyr Ile Ile Gln Lys Leu Asn Ile Val Asn Lys Lys Glu 
        115                 120                 125 

Lys Ala Thr Gly Arg Glu Val Thr His Ile Gln Phe Thr Ser Trp Pro 
    130                 135                 140 

Asp His Gly Val Pro Glu Asp Pro His Leu Leu Leu Lys Leu Arg Arg 
145                 150                 155                 160 

Arg Val Asn Ala Phe Ser Asn Phe Phe Ser Gly Pro Ile Val Val His 
                165                 170                 175 

Cys Ser Ala Gly Val Gly Arg Thr Gly Thr Tyr Ile Gly Ile Asp Ala 
            180                 185                 190 

Met Leu Glu Gly Leu Glu Ala Glu Asn Lys Val Asp Val Tyr Gly Tyr 
        195                 200                 205 

Val Val Lys Leu Arg Arg Gln Arg Cys Leu Met Val Gln Val Glu Ala 
    210                 215                 220 

Gln Tyr Ile Leu Ile His Gln Ala Leu Val Glu Tyr Asn Gln Arg Gly 
225                 230                 235                 240 

Glu Thr Glu Val Asn Leu Ser Glu 
                245 

 
           
             11  
             273  
             PRT  
             Homosapiens  
           
            11 

Asn Lys Ser Lys Asn Arg Asn Ser Asn Val Ile Pro Tyr Asp Tyr Asn 
 1               5                  10                  15 

Arg Val Pro Leu Lys His Glu Leu Glu Met Ser Lys Glu Ser Glu His 
            20                  25                  30 

Asp Ser Asp Glu Ser Ser Asp Asp Asp Ser Asp Ser Glu Glu Pro Ser 
        35                  40                  45 

Lys Tyr Ile Asn Ala Ser Phe Ile Met Ser Tyr Trp Lys Pro Glu Val 
    50                  55                  60 

Met Ile Ala Ala Gln Gly Pro Leu Lys Glu Thr Ile Gly Asp Phe Trp 
65                  70                  75                  80 

Gln Met Ile Phe Gln Arg Lys Val Lys Val Ile Val Met Leu Thr Glu 
                85                  90                  95 

Leu Lys His Gly Asp Gln Glu Ile Cys Ala Gln Tyr Trp Gly Glu Gly 
            100                 105                 110 

Lys Gln Thr Tyr Gly Asp Ile Glu Val Asp Leu Lys Asp Thr Asp Lys 
        115                 120                 125 

Ser Ser Thr Tyr Thr Leu Arg Val Phe Glu Leu Arg His Ser Lys Arg 
    130                 135                 140 

Lys Asp Ser Arg Thr Val Tyr Gln Tyr Gln Tyr Thr Asn Trp Ser Val 
145                 150                 155                 160 

Glu Gln Leu Pro Ala Glu Pro Lys Glu Leu Ile Ser Met Ile Gln Val 
                165                 170                 175 

Val Lys Gln Lys Leu Pro Gln Lys Asn Ser Ser Glu Gly Asn Lys His 
            180                 185                 190 

His Lys Ser Thr Pro Leu Leu Ile His Cys Arg Asp Gly Ser Gln Gln 
        195                 200                 205 

Thr Gly Ile Phe Cys Ala Leu Leu Asn Leu Leu Glu Ser Ala Glu Thr 
    210                 215                 220 

Glu Glu Val Val Asp Ile Phe Gln Val Val Lys Ala Leu Arg Lys Ala 
225                 230                 235                 240 

Arg Pro Gly Met Val Ser Thr Phe Glu Gln Tyr Gln Phe Leu Tyr Asp 
                245                 250                 255 

Val Ile Ala Ser Thr Tyr Pro Ala Gln Asn Gly Gln Val Lys Lys Asn 
            260                 265                 270 

Asn 

 
           
             12  
             7  
             PRT  
             Artificial Sequence  
             
               Conserved Protein Sequence  
             
           
            12 

Lys Cys Ala Gln Tyr Trp Pro 
 1               5 

 
           
             13  
             7  
             PRT  
             Artificial Sequence  
             
               Conserved Protein Sequence  
             
           
            13 

His Cys Ser Ala Gly Ile Gly 
 1               5