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

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application is a Divisional of U.S. application Ser. No. 09/191,647, filed Nov. 13, 1998 U.S. Pat. No. 6,046,015, which claims the benefit of U.S. Provisional Application No. 60/081,057 filed Apr. 07, 1998 and U.S. Provisional Application No. 60/065,544, filed Nov. 14, 1997. 
    
    
     The research carried out in the subject application was supported in part by NIH grant NS18366. The government may have rights in any patent issuing on this application. 
    
    
     INTRODUCTION 
     1. Field of the Invention 
     The field of this invention is methods for modulating nerve cell function. 
     2. Background 
     In the developing CNS, most growth cones confront the midline at one or multiple times during their journey and make the decision of whether to cross or not to cross. This decision is not a static one but rather changes according to the growth cone&#39;s history. For example, in the Drosophila ventral nerve cord, about 10% of the interneurons project their axons only on their own side, in some cases extending near the midline without crossing it. The other 90% of the interneurons first project their axons across the midline and then turn to project longitudinally on the other side, often extending near the midline. These growth cones, having crossed the midline once, never cross it again, in spite of their close proximity to the midline and the many commissural axons crossing it. This decision to cross or not to cross is not unique to Drosophila but is common to a variety of midline structures in all bilaterally symmetric nervous systems. 
     What midline signals and growth cone receptors control whether growth cones do or do not cross the midline? After crossing once, what mechanism prevents these growth cones from crossing again? A related issue concerns the nature of the midline as an intermediate target. If so many growth cones find the midline such an attractive structure, why do they cross over it rather than linger? Why do they leave the midline? 
     One approach to find the genes encoding the components of such a system is to screen for mutations in which either too many or too few axons cross the midline. Such a large-scale mutant screen was previously conducted in Drosophila, and led to the identification of two key genes:  commissureless  ( comm ) and  roundabout  ( robo ) (Seeger et al., 1993; reviewed by Tear et al., 1993). In  comm  mutant embryos, commissural growth cones initially orient toward the midline but then fail to cross it and instead recoil and extend on their own side.  robo  mutant embryos, on the other hand, display the opposite phenotype in that too many axons cross the midline; many growth cones that normally extend only on their own side instead now project across the midline and axons that normally cross the midline only once instead appear to cross and recross multiple times (Seeger et al, 1993; present disclosure). Double mutants of  comm  and  robo  display a robo-like phenotype. 
     How do  comm  and  robo  function to control midline crossing? Neither the initial paper on these genes (Seeger et al., 1993) nor the cloning of  comm  (Tear et al., 1996) resolved this question.  comm  encodes a novel surface protein expressed on midline cells. In fact, the  comm  paper (Tear et al., 1996) ended with the hope that future work would “. . . help shed some light on the enigmatic function of Comm.” 
     U.S. Ser. No. 08/971,172 ( Robo, A Novel Family of Polypeptides and Nucleic Acids , by inventors: Corey S. Goodman, Thomas Kidd, Kevin J. Mitchell and Guy Tear) discloses the cloning and characterization of  robo  in various species including Drosophila; R6bo polypeptides and polypeptide-encoding nucleic acids are also disclosed and their genbank accession numbers referenced in Kidd et al. (1998) Cell 92, 205-215.  robo  encodes a new class of guidance receptor with 5 immunoglobulin (Ig) domains, 3 fibronectin type III domains, a transmembrane domain, and a long cytoplasmic domain. Robo defines a new subfamily of Ig superfamily proteins that is highly conserved from fruit flies to mammals. The Robo ectodomains, and in particular the first two Ig domains, are highly conserved from fruit fly to human, while the cytoplasmic domains are more divergent. Nevertheless, the cytoplasmic domains contain three highly conserved short proline-rich motifs which may represent binding sites for SH3 or other binding domains in linker or signaling molecules. 
     For those axons that never cross the midline, Robo is expressed on their growth cones from the outset; for the majority of axons that do cross the midline, Robo is expressed at high levels on their growth cones only after they cross the midline. Transgenic rescue experiments in Drosophila reveal that Robo can function in a cell autonomous fashion, consistent with it functioning as a receptor. Thus, in Drosophila, Robo appears to function as the gatekeeper controlling midline crossing; growth cones expressing high levels of Robo are prevented from crossing the midline. Robo proteins in mammals function in a similar manner in controlling axon guidance. 
     U.S. Ser. No. 60/065,54 ( Methods for Modulating Nerve Cell Function , by inventors: Corey S. Goodman, Thomas Kidd, Guy Tear, Claire Russell and Kevin Mitchell, now abandoned) discloses ectopic and overexpression studies revealing that Comm down-regulates Robo expression, demonstrating that Comm functions to suppress the Robo-mediated midline repulsion. These results show that the levels of Comm at the midline and Robo on growth cones are tightly intertwined and dynamically regulated to assure that only certain growth cones cross the midline, that those growth cones that cross do not linger at the midline, and that once they cross they never do so again. 
     Relevant Literature 
     Seeger, M., Tear, G., Ferres-Marco, D. and Goodman C. S. (1993) Neuron 10, 409-426; Tear G., et al. (1996) Neuron 16, 501-514; Rothberg et al. (1990) Genes Dev 4, 2169-2187; Kidd et al. (1998) Cell 92, 205-215. 
     SUMMARY OF THE INVENTION 
     The invention provides methods and compositions relating to vertebrate Slit1 and Slit2, collectively vertebrate Slit) polypeptides, related nucleic acids, polypeptide domains thereof having vertebrate Slit-specific structure and activity, and modulators of vertebrate Slit function. Vertebrate Slit polypeptides can regulate cell, especially nerve cell, function and morphology. The polypeptides may be produced recombinantly from transformed host cells from the subject vertebrate Slit polypeptide encoding nucleic acids or purified from mammalian cells. The invention provides isolated vertebrate Slit hybridization probes and primers capable of specifically hybridizing with natural vertebrate Slit genes, vertebrate Slit-specific binding agents such as specific antibodies, and methods of making and using the subject compositions ill diagnosis (e.g. genetic hybridization screens for vertebrate Slit transcripts), therapy (e.g. to modulate nerve cell growth) and in the biopharnaceutical industry (e.g. as immunogens, reagents for isolating vertebrate Slit genes and polypeptides, reagents for screening chemical libraries for lead pharmacological agents, etc.). 
     The invention also provides methods and compositions for identifying agents which modulate the interaction of Robo and a Robo ligand and for modulating the interaction of Robo and a Robo ligand. The methods for identifying Robo:ligand modulators find particular application in commercial drug screens. These methods generally comprise (1) combining a Robo polypeptide, a Slit polypeptide and a candidate agent under conditions whereby, but for the presence of the agent, the Robo and Slit polypeptides engage in a first interaction, and (2) determining a second interaction of the Robo and Slit polypeptides in the presence of the agent, wherein a difference between the first and second interactions indicates that the agent modulates the interaction of the Robo and Slit polypeptides. The subject methods of modulating the interaction of Robo and a Robo ligand involve combining a Robo polypeptide, a Slit polypeptide and a modulator under conditions whereby, but for the presence of the modulator, the Robo and Slit polypeptides engage in a first interaction, whereby the Robo and Slit polypeptides engage in a second interaction different from the first interaction. In a particular embodiment, the modulator is dominant negative form of the Robo or Slit polypeptide. 
     DETAILED DESCRIPTION OF THE INVENTION 
     The subject methods include screens for agents which modulate Robo:ligand interactions and methods for modulating Robo:ligand interactions. Robo activation is found to regulate a wide variety of cell functions, including cell-cell interactions, cell mobility, morphology, etc. Slit polypeptides are disclosed as specific activators and inactivators of Robo polypeptides. Accordingly, the invention provides methods for modulating targeted cell function comprising the step of modulating Robo activation by contacting the cell with a modulator of a Robo:Slit interaction.. 
     The targeted Robo polypeptide is generally naturally expressed on the targeted cells. The nucleotide sequences of exemplary natural cDNAs encoding drosophila 1, drosophila 2, C. elegans, human 1, human 2 and mouse 1 Robo polypeptides and their translates are described in Kidd et al. (1998) Cell 92, 205-215 and U.S. Ser. No. 08/971,172. The targeted Robo polypeptides comprise at least a finctional Robo domain, which domain has Robo-specific amino acid sequence and binding specificity or function. Preferred Robo domains comprise at least 8, preferably at least 16, more preferably at least 32, most preferably at least 64 consecutive residues of a natural full length Robo. In a particular embodiment, the domains comprise one or more structural/functional Robo immunoglobulin, fibronectin or cytoplasmic motif domains described herein. The subject domains provide Robo-specific antigens and/or immunogens, especially when coupled to carrier proteins. For example, peptides corresponding to Robo- and human Robo-specific domains are covalently coupled to keyhole limpet antigen (KLH) and the conjugate is emulsified in Freunds complete adjuvant. Laboratory rabbits are immunized according to conventional protocol and bled. The presence of Robo-specific antibodies is assayed by solid phase immunosorbant assays using immobilized Robo polypeptides. Generic Robo-specific peptides are readily apparent as conserved regions in aligned Robo polypeptide sequences. In addition, species-specific antigenic and/or immunogenic peptides are readily apparent as diverged extracellular or cytosolic regions in alignments Human Robo-specific antibodies are characterized as uncross-reactive with non-human Robo polypeptides. 
     The subject domains provide Robo domain specific activity or function, such as Robo-specific cell, especially neuron modulating or modulating inhibitory activity, Robo-ligand-binding or binding inhibitory activity. Robo-specific activity or function may be determined by convenient in vitro, cell-based, or in vivo assays: e.g. in vitro binding assays, cell culture assays, in animals (e.g. gene therapy, transgenics, etc.), etc. The binding target may be a natural intracellular binding target, a Robo regulating protein or other regulator that directly modulates Robo activity or its localization; or non-natural binding target such as a specific immune protein such as an antibody, or a Robo specific agent such as those identified in screening assays such as described below. Robo-binding specificity may be assayed by binding equilibrium constants (usually at least about 10 7  M −1 , preferably at least about 10 8  M −1 , more preferably at least about 10 9  M −1 ), by the ability of the subject polypeptide to function as negative mutants in Robo-expressing cells, to elicit Robo specific antibody in a heterologous host (e.g a rodent or rabbit), etc. 
     Similarly, the Slit polypeptide is conveniently selected from Slit polypeptides which specifically activate or inhibit the activation of the Robo polypeptide. Exemplary suitable Slit polypeptides (a) comprises a vertebrate Slit sequence disclosed herein, especially human Slit-1 (SEQ ID NO:02), or a deletion mutant thereof which specifically modulates Robo expression or a sequence about 60-70%, preferably about 70-80%, more preferably about 80-90%, more preferably about 90-95%, most preferably about 95-99% similar to a vertebrate Slit sequence disclosed herein as determined by Best Fit analysis using default settings and is other than a natural drosophila Slit sequence, preferably other than a natural invertebrate Slit sequence, and/or (b) is encoded by a nucleic acid comprising a natural Slit encoding sequence (such as a natural human Slit-1 encoding sequence, SEQ ID NO:01) or a fragment thereof at least 36, preferably at least 72, more preferably at least 144, most preferably at least 288 nucleotides in length which specifically hybridizes thereto. Suitable deletion mutants are readily screened in Robo binding or activation assays as described herein. Preferred Slit domains/deletion mutants/fragments comprise at least 8, preferably at least 16, more preferably at least 32, most preferably at least 64 consecutive residues of a disclosed vertebrate Slit sequences and provide a Slit specific activity, such as Slit-specific antigenicity and/or immunogenicity, especially when coupled to carrier proteins as described above for Robo above. Suitable natural Slit encoding sequence fragements are of length sufficient to encode such Slit domains. In a particular embodiment, the Slit fragments comprise species specific fragments; such fragments are readily discerned from alignments of the disclosed sequences, see, e.g. shown as unboxed sequences in Tables 1 and 2. Exemplary such human Slit-1 immunogenic and/or antigenic peptides are shown in Table 3. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Alignment of human Slit-1 (SEQ ID NO:02), human Slit-2 (SEQ ID NOS:03-06), Drosphila 
               
               
                 Slit-1 (SEQ ID NO:07),  C. elegans  Slit-1 (SEQ ID NOS:08-09), mouse Slit-2 (SEQ ID NOS:10-11) 
               
               
                 and mouse Slit-1 (SEQ ID NOS:12-14). 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                  1 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  40 21 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  80 35 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  120 74 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  154 115 36 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  176 155 76 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  216 195 116 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  256 235 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  292 275 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  332 314 36 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  372 354 76 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  1 412 394 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  2 452 434 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  42 492 474 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  82 531 514 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  122 571 554 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  611 594 24 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  626 633 34 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  665 672 104 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  1 705 712 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  16 744 752 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  56 784 791 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  96 824 831 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  136 864 871 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  176 904 911 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  216 944 951 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  254 982 991 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  293 1007 1015 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit2 
               
               
                   
               
               
                  333 1047 1055 1 1 6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  373 1087 1095 24 41 46 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  412 1126 1133 62 78 80 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  451 116 1170 99 115 117 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  489 1206 1210 155 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit2 
               
               
                   
               
               
                  529 1246 1250 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  569 1284 1288 6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  609 1324 1328 46 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  617 1354 1368 86 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  653 1394 1405 123 1 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  655 1405 1445 163 25 32 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  689 1440 1485 203 64 71 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  729 1477 1523 241 102 109 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Alignment of human Slit-1 (SEQ ID NO:02), human Slit-2 (SEQ ID NOS:03-06), Drosphila 
               
               
                 Slit-1 (SEQ ID NO:07),  C. elegans  Slit-1 (SEQ ID NOS:08-09), mouse Slit-2 (SEQ ID NOS:10-11) 
               
               
                 and mouse Slit-1 (SEQ ID NOS:12-14). 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                  1 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  40 21 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  80 35 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  120 74 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  154 115 36 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  176 155 76 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  216 195 116 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  256 235 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  292 275 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  332 314 36 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  372 354 76 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  1 412 394 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  2 452 434 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  42 492 474 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  82 531 514 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  122 571 554 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  611 594 24 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  626 633 34 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  665 672 104 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  1 705 712 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  16 744 752 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  56 784 791 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  96 824 831 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  136 864 871 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  176 904 911 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  216 944 951 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  254 982 991 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  293 1007 1015 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit2 
               
               
                   
               
               
                  333 1047 1055 1 1 6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  373 1087 1095 24 41 46 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  412 1126 1133 62 78 80 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  451 116 1170 99 115 117 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  489 1206 1210 155 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit2 
               
               
                   
               
               
                  529 1246 1250 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  569 1284 1288 6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  609 1324 1328 46 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  617 1354 1368 86 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  653 1394 1405 123 1 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  655 1405 1445 163 25 32 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  689 1440 1485 203 64 71 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  729 1477 1523 241 102 109 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Alignment of human Slit-1 (SEQ ID NO:02), human Slit-2 (SEQ ID NOS:03-06), Drosphila 
               
               
                 Slit-1 (SEQ ID NO:07),  C. elegans  Slit-1 (SEQ ID NOS:08-09), mouse Slit-2 (SEQ ID NOS:10-11) 
               
               
                 and mouse Slit-1 (SEQ ID NOS:12-14). 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                  1 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  40 21 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  80 35 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  120 74 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  154 115 36 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  176 155 76 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  216 195 116 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  256 235 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 
               
               
                   
               
               
                  292 275 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  332 314 36 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  372 354 76 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  1 412 394 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  2 452 434 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  42 492 474 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  82 531 514 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  122 571 554 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  611 594 24 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  626 633 34 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  665 672 104 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 D-Slit H-Slit1 H-Slit2 
               
               
                   
               
               
                  1 705 712 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  16 744 752 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  56 784 791 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  96 824 831 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  136 864 871 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  176 904 911 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  216 944 951 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  254 982 991 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 
               
               
                   
               
               
                  293 1007 1015 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit2 
               
               
                   
               
               
                  333 1047 1055 1 1 6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  373 1087 1095 24 41 46 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  412 1126 1133 62 78 80 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  451 116 1170 99 115 117 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  489 1206 1210 155 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit2 
               
               
                   
               
               
                  529 1246 1250 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  569 1284 1288 6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  609 1324 1328 46 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  617 1354 1368 86 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 
               
               
                   
               
               
                  653 1394 1405 123 1 1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  655 1405 1445 163 25 32 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  689 1440 1485 203 64 71 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
               
                  729 1477 1523 241 102 109 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 CE-Slit D-Slit H-Slit1 M-Slit1 H-Slit2 M-Slit2 
               
               
                   
               
             
          
         
       
     
     The subject domains provide Slit domain specific activity or function, such as Slit-specific cell, especially neuron modulating or modulating inhibitory activity, Slit-ligand-binding or binding inhibitory activity. Slit-specific activity or function may be determined by convenient in vitro, cell-based, or in vivo assays: e.g. in vitro binding assays, cell culture assays, in animals (e.g. gene therapy, transgenics, etc.), etc. The binding target may be a natural intracellular binding target, a Slit regulating protein or other regulator that directly modulates Slit activity or its localization; or non-natural binding target such as a specific immune protein such as an antibody, or a Slit specific agent such as those identified in screening assays such as described below. Slit-binding specificity may be assayed by binding equilibrium constants (usually at least about 10 7  M −1 , preferably at least about 10 8  M −1 , more preferably at least about 10 9  M −1 ), by the ability of the subject polypeptide to function as negative mutants in Slit-expressing cells, to elicit Slit specific antibody in a heterologous host (e.g a rodent or rabbit), etc. 
     In one embodiment, the Slit polypeptides are encoded by a nucleic acid comprising SEQ ID NO:01 or a fragment thereof which hybridizes with a full-length strand thereof, preferably under stringent conditions. Such nucleic acids comprise at least 36, preferably at least 72, more preferably at least 144 and most preferably at least 288 nucleotides of SEQ ID NO:01. Demonstrating specific hybridization generally requires stringent conditions, for example, hybridizing in a buffer comprising 30% formamide in 5×SSPE (0.18 M NaCl, 0.01 M NaPO 4 , pH7.7, 0.001 M EDTA) buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE (Conditions I); preferably hybridizing in a buffer comprising 50% formamide in 5×SSPE buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE buffer at 42° C. (Conditions II). Exemplary nucleic acids which hybridize with a strand of SEQ ID NO:01 are shown in Table 4. 
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Exemplary nucleic acids which hybridize with a strand of SEQ ID NO: 01 under 
               
               
                 Conditions I and/or II. 
               
             
          
           
               
                 Slit Nucleic Acid 
                 Hybridization 
                 Slit Nucleic Acid 
                 Hybridization 
               
               
                   
               
               
                 SEQ ID NO: 01, nucl. 1-47 
                 + 
                 SEQ ID NO: 01, nucl. 1258-1279 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 58-99 
                 + 
                 SEQ ID NO: 01, nucl. 1375-1389 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 95-138 
                 + 
                 SEQ ID NO: 01, nucl. 1581-1595 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 181-220 
                 + 
                 SEQ ID NO: 01, nucl. 1621-1639 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 261-299 
                 + 
                 SEQ ID NO: 01, nucl. 1744-1755 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 274-315 
                 + 
                 SEQ ID NO: 01, nucl. 1951-1969 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 351-389 
                 + 
                 SEQ ID NO: 01, nucl. 2150-2163 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 450-593 
                 + 
                 SEQ ID NO: 01, nucl. 2524-2546 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 524-546 
                 + 
                 SEQ ID NO: 01, nucl: 2761-2780 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 561-608 
                 + 
                 SEQ ID NO: 01, nucl. 2989-2999 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 689-727 
                 + 
                 SEQ ID NO: 01, nucl. 3108-3117 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 708-737 
                 + 
                 SEQ ID NO: 01, nucl. 3338-3351 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 738-801 
                 + 
                 SEQ ID NO: 01, nucl. 3505-3514 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 805-854 
                 + 
                 SEQ ID NO: 01, nucl. 3855-3867 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 855-907 
                 + 
                 SEQ ID NO: 01, nucl. 4010-4025 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 910-953 
                 + 
                 SEQ ID NO: 01, nucl. 4207-4219 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 1007-1059 
                 + 
                 SEQ ID NO: 01, nucl. 4333-4345 
                 + 
               
               
                 SEQ ID NO: 01, nucl. 1147-1163 
                 + 
                 SEQ ID NO: 01, nucl. 4521-4529 
                 + 
               
               
                   
               
             
          
         
       
     
     A wide variety of cell types express Robo polypeptides subject to regulation by the disclosed methods, including many neuronal cells, transformed cells, infected (e.g. virus) cells, etc. Ascertaining Robo binding or activation is readily effected by binding assays or cells function assays as disclosed herein or in the cited copending applications. Accordingly, indications for the subject methods encompass a wide variety of cell types and function, including axon outgrowth, tumor cell invasion or migration, etc. The target cell may reside in culture or in situ, i.e. within the natural host. For in situ applications, the compositions are added to a retained physiological fluid such as blood or synovial fluid. For CNS administration, a variety of techniques are available for promoting transfer of the therapeutic across the blood brain barrier including disruption by surgery or injection, drugs which transiently open adhesion contact between CNS vasculature endothelial cells, and compounds which facilitate translocation through such cells. Slit polypeptides may also be amenable to direct injection or infusion, topical, intratracheal/nasal administration e.g. through aerosol, intraocularly, or within/on implants e.g. fibers e.g. collagen, osmotic pumps, grafts comprising appropriately transformed cells, etc. A particular method of administration involves coating, embedding or derivatizing fibers, such as collagen fibers, protein polymers, etc. with therapeutic polypeptides. Other useful approaches are described in Otto et al. (1989) J Neuroscience Research 22, 83-91 and Otto and Unsicker (1990) J Neuroscience 10, 1912-1921. Generally, the amount administered will be empirically determined, typically in the range of about 10 to 1000 μg/kg of the recipient and the concentration will generally be in the range of about 50 to 500 μg/ml in the dose administered. Other additives may be included, such as stabilizers, bactericides, etc. will be present in conventional amounts. 
     In one embodiment, the invention provides administering the subject Slit polypeptides in combination with a pharmaceutically acceptable excipient such as sterile saline or other medium, gelatin, an oil, etc. to form pharmaceutically acceptable compositions. The compositions and/or compounds may be administered alone or in combination with any convenient carrier, diluent, etc. and such administration may be provided in single or multiple dosages. Useful carriers include solid, semi-solid or liquid media including water and non-toxic organic solvents. In another embodiment, the invention provides the subject compounds in the form of a pro-drug, which can be metabolically converted to the subject compound by the recipient host. A wide variety of prodrug formulations for polypeptide-based therapeutics are known in the art. The compositions may be provided in any convenient form including tablets, capsules, troches, powders, sprays, creams, etc. As such the compositions, in pharmaceutically acceptable dosage units or in bulk, may be iincorporated into a wide variety of containers. For example, dosage units may be included in a variety of containers including capsules, pills, etc. The compositions may be advantageously combined and/or used in combination with other therapeutic or prophylactic agents, different from the subject compounds. In many instances, administration in conjunction with the subject compositions enhances the efficacy of such agents, see e.g.  Goodman &amp; Gilman&#39;s The Pharmacological Basis of Therapeutics,  9 th  Ed., 1996, McGraw-Hill. 
     In another aspect, the invention provides methods of screening for agents which modulate Robo-Slit interactions. These methods generally involve forming a mixture of a Robo-expressing cell, a Slit polypeptide and a candidate agent, and determining the effect of the agent on the amount of Robo expressed by the cell. The methods are amenable to automated, cost-effective high throughput screening of chemical libraries for lead compounds. Identified reagents find use in the pharmaceutical industries for animal and human trials; for example, the reagents may be derivatized and rescreened in in vitro and in vivo assays to optimize activity and minimize toxicity for pharmaceutical development. Cell and animal based neural guidance/repulsion assays are described in detail in the experimental section below. 
     The amino acid sequences of the disclosed vertebrate Slit polypeptides are used to back-translate Slit polypeptide-encoding nucleic acids optimized for selected expression systems (Holler et al. (1993) Gene 136, 323-328; Martin et al. (1995) Gene 154, 150-166) or used to generate degenerate oligonucleotide primers and probes for use in the isolation of natural Slit-encoding nucleic acid sequences (“GCG” software, Genetics Computer Group, Inc, Madison Wis.). Slit-encoding nucleic acids used in Slit-expression vectors and incorporated into recombinant host cells, e.g. for expression and screening, transgenic animals, e.g. for finctional studies such as the efficacy of candidate drugs for disease associated with Slit-modulated cell function, etc. 
     The invention also provides nucleic acid hybridization probes and replication/amplification primers having a vertebrate Slit cDNA specific sequence comprising a fragment of a disclosed vertebrate cDNA sequence, and sufficient to effect specific hybridization thereto. Such primers or probes are at least 12, preferably at least 24, more preferably at least 36 and most preferably at least 96 nucleotides in length. Demonstrating specific hybridization generally requires stringent conditions, for example, hybridizing in a buffer comprising 30% formamide in5×SSPE (0.18 M NaCl, 0.01 M NaPO 4 , pH7.7, 0.001 M EDTA) buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE; preferably hybridizing in a buffer comprising 50% formamide in 5×SSPE buffer at a temperature of 42° C. and remaining bound when subject to washing at 42° C. with 0.2×SSPE buffer at 42° C. Slit nucleic acids can also be distinguished using alignment algorithms, such as BLASIX (Altschul et al. (1990) Basic Local Alignment Search Tool, J Mol Biol 215, 403-410). In addition, the invention provides nucleic acids having a sequence about 60-70%, preferably about 70-80%, more preferably about 80-90%, more preferably about 90-95%, most preferably about 95-99% similar to a vertebrate Slit sequence disclosed herein as determined by Best Fit analysis using default settings and is other than a natural drosophila Slit sequence, preferably other than a natural invertebrate Slit sequence. In a particular embodiment, the Slit polynucleotide fragments comprise species specific fragments; such fragments are readily discerned from alignments of the disclosed sequences. 
     The subject nucleic acids are of synthetic/non-natural sequences and/or are recombinant, meaning they comprise a non-natural sequence or a natural sequence joined to nucleotide(s) other than that which it is joined to on a natural chromosome. The subject recombinant nucleic acids comprising the nucleotide sequence of disclosed vertebrate Slit nucleic acids, or fragments thereof, contain such sequence or fragment at a terminus, immediately flanked by (i.e. contiguous with) a sequence other than that which it is joined to on a natural chromosome, or flanked by a native flanking region fewer than 10 kb, preferably fewer than 2 kb, more preferably fewer than 500 bp, which is at a terminus or is immediately flanked by a sequence other than that which it is joined to on a natural chromosome. While the nucleic acids are usually RNA or DNA, it is often advantageous to use nucleic acids comprising other bases or nucleotide analogs to provide modified stability, etc. 
     The subject nucleic acids find a wide variety of applications including use as translatable transcripts, hybridization probes, PCR primers, diagnostic nucleic acids, etc.; use in detecting the presence of Slit genes and gene transcripts and in detecting or amplifying nucleic acids encoding additional Slit homologs and structural analogs. In diagnosis, Slit hybridization probes find use in identifying wild-type and mutant Slit alleles in clinical and laboratory samples. Mutant alleles are used to generate allele-specific oligonucleotide (ASO) probes for high-throughput clinical diagnoses. In therapy, therapeutic Slit nucleic acids are used to modulate cellular expression or intracellular concentration or availability of active Slit. Exemplary humain Slit-1 probes and primers are shown in Table 5 and Table 6. 
     
       
         
               
             
               
               
             
           
               
                 TABLE 5 
               
               
                   
               
               
                 Hybridization Probes for Regions of Human Slit-1. 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Hybridization probe for first leucine rich 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat region 
                 82-828 
               
               
                 Hybridization probe for second leucine 
                 SEQ ID NO: 01, nucleotides 
               
               
                 rich repeat region 
                 829-1503 
               
               
                 Hybridization probe for third leucine rich 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat region 
                 1504-2166 
               
               
                 Hybridization probe for fourth leucine rich 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat region 
                 2167-2751 
               
               
                 Hybridization probe for EGF repeats one 
                 SEQ ID NO: 01, nucleotides 
               
               
                 to five 
                 2752-3327 
               
               
                 Hybridization probe for the sixth EGF 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat and preceding spacer region 
                 3328-3461 
               
               
                 Hybridization probe for the 99aa spacer/G- 
                 SEQ ID NO: 01, nucleotides 
               
               
                 loop region 
                 3462-3987 
               
               
                 Hybridization probe for EGF repeats seven 
                 SEQ ID NO: 01, nucleotides 
               
               
                 to nine 
                 3988-4341 
               
               
                 Hybridization probe for the cysteine knot 
                 SEQ ID NO: 01, nucleotides 
               
               
                 region 
                 4342-4575 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 5 
               
               
                   
               
               
                 Hybridization Probes for Regions of Human Slit-1. 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Hybridization probe for first leucine rich 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat region 
                 82-828 
               
               
                 Hybridization probe for second leucine 
                 SEQ ID NO: 01, nucleotides 
               
               
                 rich repeat region 
                 829-1503 
               
               
                 Hybridization probe for third leucine rich 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat region 
                 1504-2166 
               
               
                 Hybridization probe for fourth leucine rich 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat region 
                 2167-2751 
               
               
                 Hybridization probe for EGF repeats one 
                 SEQ ID NO: 01, nucleotides 
               
               
                 to five 
                 2752-3327 
               
               
                 Hybridization probe for the sixth EGF 
                 SEQ ID NO: 01, nucleotides 
               
               
                 repeat and preceding spacer region 
                 3328-3461 
               
               
                 Hybridization probe for the 99aa spacer/G- 
                 SEQ ID NO: 01, nucleotides 
               
               
                 loop region 
                 3462-3987 
               
               
                 Hybridization probe for EGF repeats seven 
                 SEQ ID NO: 01, nucleotides 
               
               
                 to nine 
                 3988-4341 
               
               
                 Hybridization probe for the cysteine knot 
                 SEQ ID NO: 01, nucleotides 
               
               
                 region 
                 4342-4575 
               
               
                   
               
             
          
         
       
     
     Leucine rich repeats (LRRs) are predicted by comparison with known proteins and by the presence of a leucine rich core sequence. In slit proteins, the LRRs are flanked by conserved sequences referred to as the amino- and carboxy- flanking regions. These flanking regions are found in other known proteins, but only in a few instances are both the amino- and carboxy- flank regions present in a single protein. The so called “99aa spacer.” is actually ˜200 amino acids in the Drosophila protein and 174 amino acids in Human Slit-1. This region shows homology to the G-loops of laminin A chains. 
     Cysteine knots are dimerisation domains defined by the presence of six cysteine residues between which disulphide bridges form. The only absolutely conserved residues are the six cysteines, and spacing between them is highly variable, apart from between cysteines 2 and 3, and 5 and 6. The glycine between cysteines 2 and 3 is only present in a subset of cysteine knots. Drosophila slit and Human slit-1 both have an extra cysteine after cysteines 5 and 6: this may serve as an intermolecular bond. Human Slit-1 gene displays the overall structure of the Drosophila gene, and amino acid conservation is found along the entire length of the protein (48% homology at the amino acid sequence excluding the signal sequence; see below). The Human gene has an extra LRR between LRR2 and LRR3 of the first set of LRRs; in the third set, the Human gene has an extra LRR between LRR3 and LRR4. The Human gene has two extra EGF repeats, on either side of the seventh EGF repeat in Drosophila slit. 
     Isolation of Human slit-1 
     Searching of the EST database revealed an EST, ab16g10.rl1, with homology to the 99aa spacer region of Drosophila slit. This EST was used to probe a Human fetal brain library (Stratagene), and clones for Human slit-1 were isolated. 
     
       
         
               
             
               
               
             
               
             
               
               
             
           
               
                   
               
             
             
               
                 Features of Human Slit Predicted Protein 
               
               
                   
               
             
          
           
               
                 Signal sequence 
                 SEQ ID NO: 02, residues 7-24 
               
               
                 First amino-flanking sequence 
                 SEQ ID NO: 02, residues 28-59 
               
               
                 First set of Leucine Rich Repeats 
                 SEQ ID NO: 02, residues 60-179 (6 repeats) 
               
               
                 First carboxy-flanking sequence 
                 SEQ ID NO: 02, residues 180-276 
               
               
                 Second amino-flanking sequence 
                 SEQ ID NO: 02, residues 277-308 
               
               
                 Second set of Leucine Rich Repeats 
                 SEQ ID NO: 02, residues 309-434 (5 repeats) 
               
               
                 Second carboxy-flanking sequence 
                 SEQ ID NO: 02, residues 435-501 
               
               
                 Third amino-flanking sequence 
                 SEQ ID NO: 02, residues 502-533 
               
               
                 Third set of Leucine Rich Repeats 
                 SEQ ID NO: 02, residues 534-560 (5 repeats) 
               
               
                 Third carboxy-flanking sequence 
                 SEQ ID NO: 02, residues 661-722 
               
               
                 Fourth amino-flanking sequence 
                 SEQ ID NO: 02, residues 723-754 
               
               
                 Fourth set of Leucine Rich Repeats 
                 SEQ ID NO: 02, residues 755-855 (4 repeats) 
               
               
                 Fourth carboxy-flanking sequence 
                 SEQ ID NO: 02, residues 856-917 
               
               
                 First EGF repeat 
                 SEQ ID NO: 02, residues 918-952 
               
               
                 Second EGF repeat 
                 SEQ ID NO: 02, residues 953-993 
               
               
                 Third EGF repeat 
                 SEQ ID NO: 02, residues 994-1031 
               
               
                 Fourth EGF repeat 
                 SEQ ID NO: 02, residues 1032-1071 
               
               
                 Fifth EGF repeat 
                 SEQ ID NO: 02, residues 1072-1109 
               
               
                 Spacer 
                 SEQ ID NO: 02, residues 1110-1116 
               
               
                 Sixth EGF repeat 
                 SEQ ID NO: 02, residues 1117-1153 
               
               
                 “99aa spacer” 
                 SEQ ID NO: 02, residues 1155-1329 
               
               
                 Seventh EGF repeat 
                 SEQ ID NO: 02, residues 1330-1366 
               
               
                 Eighth EGF repeat 
                 SEQ ID NO: 02, residues 1367-1404 
               
               
                 Nineth EGF repeat 
                 SEQ ID NO: 02, residues 1405-1447 
               
               
                 Cysteine knot motif 
                 SEQ ID NO: 02, residues 1448-1525 
               
             
          
           
               
                 Amino acid identity between Drosophila and Human Slit-1 
               
               
                   
               
             
          
           
               
                 First amino-flanking sequence 
                 53% 
               
               
                 First set of Leucine Rich Repeats 
                 52% (54%, 67%, NA, 38%, 54%, 50%) 
               
               
                 First carboxy-flanking sequence 
                 42% 
               
               
                 Second amino-flanking sequence 
                 50% 
               
               
                 Second set of Leucine Rich Repeats 
                 60% (54%, 58%, 67%, 71%, 50%) 
               
               
                 Second carboxy-flanking sequence 
                 62% 
               
               
                 Third amino-flanking sequence 
                 56% 
               
               
                 Third set of Leucine Rich Repeats 
                 49% (46%, 46%, 42%, NA, 58%) 
               
               
                 Third carboxy-flanking sequence 
                 36% 
               
               
                 Fourth amino-flanking sequence 
                 53% 
               
               
                 Fourth set of Leucine Rich Repeats 
                 48% (25%, 58%, 46%, 63%) 
               
               
                 Fourth carboxy-flanking sequence 
                 63% 
               
               
                 First EGF repeat 
                 34% 
               
               
                 Second EGF repeat 
                 46% 
               
               
                 Third EGF repeat 
                 46% 
               
               
                 Fourth EGF repeat 
                 35% 
               
               
                 Fifth EGF repeat 
                 47% 
               
               
                 Spacer 
                 22% 
               
               
                 Sixth EGF repeat 
                 40% 
               
               
                 “99aa spacer” 
                 38% 
               
               
                 Seventh EGF repeat 
                 11%/NA 
               
               
                 Eighth EGF repeat 
                 44% 
               
               
                 Nineth EGF repeat 
                 29%/NA 
               
               
                 Cysteine knot motif 
                 34% 
               
               
                   
               
               
                 NA: not applicable due to absence of homologous repeat.  
               
               
                 Figures for individual LLRs are shown in brackets.  
               
             
          
         
       
     
     The following examplary assay is offered by way of illustration and not by way of limitation: 
    
    
     EXAMPLES 
     Protocol for Ligand Screening of Transfected COS cells. 
     I. Prepare the Ligand 
     Expression Construct: cDNAs encoding targeted Slit polypeptides are tagged with the Fc portion of human IgG and subcloned into a 293 expression vector (pCEP4: In Vitrogen). 
     Transfection: 293 EBNA cells are transfected (CaPO 4  method) with the Slit expression constructs. After 24 h recovery, transfected cells are selected with G418 (geneticin, 250 ug/ml, Gibco) and hygromycin (200 ug/ml). Once the selection process is complete, cells are maintained in Dulbecco&#39;s Modified Eagles medium (DME)/10% FCS under selection. 
     Preparation of Conditioned Medium: Serum-containing media is replaced with Optimem with glutamax-1 (Gibco) and 300 ng/ml heparin (Sigma), and the cells are conditioned for 3 days. The media is collected and spun at 3,000xg for 10 minutes. The supernatant is filtered (0.45 um) and stored with 0.1% azide at 4° C. for no more than 2 weeks. 
     II. Prepare Truncated Receptor (Positive Control) 
     Expression Construct: cDNA encoding a corresponding Robo C-terminal deletion mutant comprising the extracellular domain (truncated immediately N-terminal to the transmembrane region) is subdloned into a 293 expression vector (pCEP4: In Vitrogen). 
     Transfection: 293 EBNA cells are transfected (CaPO 4  method) with the receptor mutant expression construct. After 24 h recovery, transfected cells are selected with G418 (geneticin, 250 ug/ml, Gibco) and hygromycin (200 ug/ml). Once the selection process is complete, cells are maintained in Dulbecco&#39;s Modified Eagles medium (DME)/10% FCS under selection. 
     Preparation of Conditioned Medium: Serumn-containing media is replaced with Optimem with glutamax-1 (Gibco) and 300 ng/ml heparin (Sigma), and the cells are conditioned for 3 days. The media is collected and spun at 3,000xg for 10 minutes. The supernatant is filtered (0.45 um) and stored with 0.1% azide at 4° C. for no more than 2 weeks. 
     II. Transfect COS Cells 
     Seed COS cells (250,000) on 35 mm dishes in 2 ml DME/10% FCS. 
     18-24 h later, dilute 1 ug of Robo-encoding DNA (cDNA cloned into pMT21 expression vector) into 200 ul serum-free media and add 6 ul of Lipofectamine (Gibco). Incubate this solution at room temperature for 15-45 min. 
     Wash the cells 2X with PBS. Add 800 ul serum-free media to the tube containing the lipid-DNA complexes. Overlay this solution onto the washed cells. 
     Incubate for 6 h. Stop the reaction by adding 1 ml DMA/20% FCS. Refeed cells. Assay cells 12 hr later. 
     III. Ligand Binding Assay 
     Wash plates of transfected COS cells 1X with cold PBS (plus Ca/Mg)/1% goat serum. Add 1 ml conditioned media neat and incubate 90 min at room temp. 
     Wash plates 3X with PBS (plus Ca/Mg). On the 4th wash, add 1 ml 50% methanol to 1 ml PBS. Then add 1 ml methanol. Evacuate and add 1 ml methanol. 
     Wash 1X with PBS. Wash 1X PBS/1% goat serum. 
     Add secondary antibody (1-to-2,000 anti-human Fc conjugated to alkaline phosphatase (Jackson Lab)) in PBS/1% goat serum. Incubate 30-40 min room temp. 
     Wash 3X with PBS. Wash 1X alkaline phosphatase buffer (100 mM Tris-Cl, pH 9.5, 100 mM NaCl, 5 MM MgCl 2 ). Prepare alkaline phosphatase reagents: 4.5 ul/ml NBT and 3.5 ul/ml BCIP (Gibco) in alkaline phosphatase buffer. 
     Incubate 10-30 min, quench with 20 mM EDTA in PBS. Cells that have bound Slit polypeptides are visible by the presence of a dark purple reaction product. 
     In parallel incubations, positive controls are provided by titrating Slit binding with serial dilutions of the mutant receptor conditioned medium. 
     IV. Results: Binding of Slit to Robo 
     Cell expressing mammalian Slit polypeptides were shown to bind Robo. No reactivity was observed with control COS cells or with receptor-expressing COS cells in the presence of the secondary antibody but in the absence of the Slit-Fc fusion. Binding was observed to receptor-expression cells using a construct in which a Slit polypeptide is fused directly to alkaline phosphatase, for which a secondary antibody is not required. Receptor deletion imutants titrate the Slit-Robo binding, serving as a positive control for inhibition assays. 
     Protocol for high throughiut Robo-Slit binding assay. 
     A. Reagents: 
     Neutralite Avidin: 20 μg/ml in PBS. 
     Blocking buffer: 5% BSA, 0.5% Tween 20 in PBS; 1 hour at room temperature. 
     Assay Buffer: 100 mM KCl, 20 mM HEPES pH 7.6, 1 mM MgCl 2 , 1% glycerol, 0.5% NP-40, 50 mM β-mercaptoethanol, 1 mg/ml BSA, cocktail of protease inhibitors. 
         33   P Robo polypeptide 10x stock: 10 −8 -10 −6  M “cold” Robo polypeptide specific Robo domain supplemented with 200,000-250,000 cpm of labeled Robo (Beckman counter). Place in the 4° C. microfridge during screening. 
     Protease inhibitor cocktail (1000X): 10 mg Trypsin Inhibitor (BMB # 109894), 10 mg Aprotinin (BMB # 236624), 25 mg Benzamidine (Sigma # B-6506), 25 mg Leupeptin (BMB # 1017128), 10 mg APMSF (BMB # 917575), and 2mM NaVO 3  (Sigma # S-6508) in 10 ml of PBS. 
     Slit: 10 −7 -10 −5  M biotinylated Slit in PBS. 
     B. Preparation of assay plates: 
     Coat with 120 μl of stock N-Avidin per well overnight at 4° C. 
     Wash 2 times with 200 μl PBS. 
     Block with 150 μl of blocking buffer. 
     Wash 2 times with 200 μl PBS. 
     C. Assay: 
     Add 40 μl assay buffer/well. 
     Add 10 μl compound or extract. 
     Add 10 μl  33 P-Robo (20-25,000 cpm/0.1-10 pmoles/well=10 −9 -10 −7  M final conc). 
     Shake at 25° C. for 15 minutes. 
     Incubate additional 45 minutes at 25° C. 
     Add 40 μM biotinylated Slit (0.1-10 pmoles/40 ul in assay buffer) 
     Incubate 1 hour at room temperature. 
     Stop the reaction by washing 4 times with 200 μM PBS. 
     Add 150 μM scintillation cocktail. 
     Count in Topcount. 
     D. Controls for all assays (located on each plate): 
     a. Non-specific binding 
     b. Soluble (non-biotinylated Slit) at 80% inhibition. 
     All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims. 
     
       
         
           
             20 
           
           
             1 
             4758 
             DNA 
             human 
             
               CDS 
               (1)..(4575) 
             
           
            1
atg cgc ggc gtt ggc tgg cag atg ctg tcc ctg tcg ctg ggg tta gtg       48
Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val
  1               5                  10                  15
ctg gcg atc ctg aac aag gtg gca ccg cag gcg tgc ccg gcg cag tgc       96
Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys
             20                  25                  30
tct tgc tcg ggc agc aca gtg gac tgt cac ggg ctg gcg ctg cgc agc      144
Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser
         35                  40                  45
gtg ccc agg aat atc ccc cgc aac acc gag aga ctg gat tta aat gga      192
Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly
     50                  55                  60
aat aac atc aca aga att acg aag aca gat ttt gct ggt ctt aga cat      240
Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His
 65                  70                  75                  80
cta aga gtt ctt cag ctt atg gag aat aag att agc acc att gaa aga      288
Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg
                 85                  90                  95
gga gca ttc cag gat ctt aaa gaa cta gag aga ctg cgt tta aac aga      336
Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg
            100                 105                 110
aat cac ctt cag ctg ttt cct gag ttg ctg ttt ctt ggg act gcg aag      384
Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys
        115                 120                 125
cta tac agg ctt gat ctc agt gaa aac caa att cag gca atc cca agg      432
Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg
    130                 135                 140
aaa gct ttc cgt ggg gca gtt gac ata aaa aat ttg caa ctg gat tac      480
Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr
145                 150                 155                 160
aac cag atc agc tgt att gaa gat ggg gca ttc agg gct ctc cgg gac      528
Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp
                165                 170                 175
ctg gaa gtg ctc act ctc aac aat aac aac att act aga ctt tct gtg      576
Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val
            180                 185                 190
gca agt ttc aac cat atg cct aaa ctt agg act ttt cga ctg cat tca      624
Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser
        195                 200                 205
aac aac ctg tat tgt gac tgc cac ctg gcc tgg ctc tcc gac tgg ctt      672
Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu
    210                 215                 220
cgc aaa agg cct cgg gtt ggt ctg tac act cag tgt atg ggc ccc tcc      720
Arg Lys Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser
225                 230                 235                 240
cac ctg aga ggc cat aat gta gcc gag gtt caa aaa cga gaa ttt gtc      768
His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val
                245                 250                 255
tgc agt gat gag gaa gaa ggt cac cag tca ttt atg gct cct tct tgt      816
Cys Ser Asp Glu Glu Glu Gly His Gln Ser Phe Met Ala Pro Ser Cys
            260                 265                 270
agt gtt ttg cac tgc cct gcc gcc tgt acc tgt agc aac aat atc gta      864
Ser Val Leu His Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val
        275                 280                 285
gac tgt cgt ggg aaa ggt ctc act gag atc ccc aca aat ctt cca gag      912
Asp Cys Arg Gly Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu
    290                 295                 300
acc atc aca gaa ata cgt ttg gaa cag aac aca atc aaa gtc atc cct      960
Thr Ile Thr Glu Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro
305                 310                 315                 320
cct gga gct ttc tca cca tat aaa aag ctt aga cga att gac ctg agc     1008
Pro Gly Ala Phe Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser
                325                 330                 335
aat aat cag atc tct gaa ctt gca cca gat gct ttc caa gga cta cgc     1056
Asn Asn Gln Ile Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg
            340                 345                 350
tct ctg aat tca ctt gtc ctc tat gga aat aaa atc aca gaa ctc ccc     1104
Ser Leu Asn Ser Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro
        355                 360                 365
aaa agt tta ttt gaa gga ctg ttt tcc tta cag ctc cta tta ttg aat     1152
Lys Ser Leu Phe Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn
    370                 375                 380
gcc aac aag ata aac tgc ctt cgg gta gat gct ttt cag gat ctc cac     1200
Ala Asn Lys Ile Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His
385                 390                 395                 400
aac ttg aac ctt ctc tcc cta tat gac aac aag ctt cag acc atc gcc     1248
Asn Leu Asn Leu Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala
                405                 410                 415
aag ggg acc ttt tca cct ctt cgg gcc att caa act atg cat ttg gcc     1296
Lys Gly Thr Phe Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala
            420                 425                 430
cag aac ccc ttt att tgt gac tgc cat ctc aag tgg cta gcg gat tat     1344
Gln Asn Pro Phe Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr
        435                 440                 445
ctc cat acc aac ccg att gag acc agt ggt gcc cgt tgc acc agc ccc     1392
Leu His Thr Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro
    450                 455                 460
cgc cgc ctg gca aac aaa aga att gga cag atc aaa agc aag aaa ttc     1440
Arg Arg Leu Ala Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe
465                 470                 475                 480
cgt tgt tca ggt aca gaa gat tat cga tca aaa tta agt gga gac tgc     1488
Arg Cys Ser Gly Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys
                485                 490                 495
ttt gcg gat ctg gct tgc cct gaa aag tgt cgc tgt gaa gga acc aca     1536
Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr
            500                 505                 510
gta gat tgc tct aat caa aag ctc aac aaa atc ccg gag cac att ccc     1584
Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro
        515                 520                 525
cag tac act gca gag ttg cgt ctc aat aat aat gaa ttt acc gtg ttg     1632
Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu
    530                 535                 540
gaa gcc aca gga atc ttt aag aaa ctt cct caa tta cgt aaa ata aac     1680
Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn
545                 550                 555                 560
ttt agc aac aat aag atc aca gat att gag gag gga gca ttt gaa gga     1728
Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly
                565                 570                 575
gca tct ggt gta aat gaa ata ctt ctt acg agt aat cgt ttg gaa aat     1776
Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn
            580                 585                 590
gtg cag cat aag atg ttc aag gga ttg gaa agc ctc aaa act ttg atg     1824
Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met
        595                 600                 605
ttg aga agc aat cga ata acc tgt gtg ggg aat gac agt ttc ata gga     1872
Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly
    610                 615                 620
ctc agt tct gtg cgt ttg ctt tct ttg tat gat aat caa att act aca     1920
Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr
625                 630                 635                 640
gtt gca cca ggg gca ttt gat act ctc cat tct tta tct act cta aac     1968
Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn
                645                 650                 655
ctc ttg gcc aat cct ttt aac tgt aac tgc tac ctg gct tgg ttg gga     2016
Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly
            660                 665                 670
gag tgg ctg aga aag aag aga att gtc acg gga aat cct aga tgt caa     2064
Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln
        675                 680                 685
aaa cca tac ttc ctg aaa gaa ata ccc atc cag gat gtg gcc att cag     2112
Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln
    690                 695                 700
gac ttc act tgt gat gac gga aat gat gac aat agt tgc tcc cca ctt     2160
Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu
705                 710                 715                 720
tct cgc tgt cct act gaa tgt act tgc ttg gat aca gtc gtc cga tgt     2208
Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys
                725                 730                 735
agc aac aag ggt ttg aag gtc ttg ccg aaa ggt att cca aga gat gtc     2256
Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val
            740                 745                 750
aca gag ttg tat ctg gat gga aac caa ttt aca ctg gtt ccc aag gaa     2304
Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu
        755                 760                 765
ctc tcc aac tac aaa cat tta aca ctt ata gac tta agt aac aac aga     2352
Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg
    770                 775                 780
ata agc acg ctt tct aat cag agc ttc agc aac atg acc cag ctc ctc     2400
Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu
785                 790                 795                 800
acc tta att ctt agt tac aac cgt ctg aga tgt att cct cct cgc acc     2448
Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr
                805                 810                 815
ttt gat gga tta aag tct ctt cga tta ctt tct cta cat gga aat gac     2496
Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp
            820                 825                 830
att tct gtt gtg cct gaa ggt gct ttc aat gat ctt tct gca tta tca     2544
Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser
        835                 840                 845
cat cta gca att gga gcc aac cct ctt tac tgt gat tgt aac atg cag     2592
His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln
    850                 855                 860
tgg tta tcc gac tgg gtg aag tcg gaa tat aag gag cct gga att gct     2640
Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala
865                 870                 875                 880
cgt tgt gct ggt cct gga gaa atg gca gat aaa ctt tta ctc aca act     2688
Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr
                885                 890                 895
ccc tcc aaa aaa ttt acc tgt caa ggt cct gtg gat gtc aat att cta     2736
Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu
            900                 905                 910
gct aag tgt aac ccc tgc cta tca aat ccg tgt aaa aat gat ggc aca     2784
Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr
        915                 920                 925
tgt aat agt gat cca gtt gac ttt tac cga tgc acc tgt cca tat ggt     2832
Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly
    930                 935                 940
ttc aag ggg cag gac tgt gat gtc cca att cat gcc tgc atc agt aac     2880
Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn
945                 950                 955                 960
cca tgt aaa cat gga gga act tgc cac tta aag gaa gga gaa gaa gat     2928
Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp
                965                 970                 975
gga ttc tgg tgt att tgt gct gat gga ttt gaa gga gaa aat tgt gaa     2976
Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu
            980                 985                 990
gtc aac gtt gat gat tgt gaa gat aat gac tgt gaa aat aat tct aca     3024
Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr
        995                1000                1005
tgt gtc gat ggc att aat aac tac aca tgc ctt tgc cca cct gag tat     3072
Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr
   1010                1015                1020
aca ggt gag ttg tgt gag gag aag ctg gac ttc tgt gcc cag gac ctg     3120
Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu
1025               1030                1035                1040
aac ccc tgc cag cac gat tca aag tgc atc cta act cca aag gga ttc     3168
Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe
               1045                1050                1055
aaa tgt gac tgc aca cca ggg tac gta ggt gaa cac tgc gac atc gat     3216
Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp
           1060                1065                1070
ttt gac gac tgc caa gac aac aag tgt aaa aac gga gcc cac tgc aca     3264
Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr
       1075                1080                1085
gat gca gtg aac ggc tat acg tgc ata tgc ccc gaa ggt tac agt ggc     3312
Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly
   1090                1095                1100
ttg ttc tgt gag ttt tct cca ccc atg gtc ctc cct cgt acc agc ccc     3360
Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro
1105               1110                1115                1120
tgt gat aat ttt gat tgt cag aat gga gct cag tgt atc gtc aga ata     3408
Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile
               1125                1130                1135
aat gag cca ata tgt cag tgt ttg cct ggc tat cag gga gaa aag tgt     3456
Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys
           1140                1145                1150
gaa aaa ttg gtt agt gtg aat ttt ata aac aaa gag tct tat ctt cag     3504
Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln
       1155                1160                1165
att cct tca gcc aag gtt cgg cct cag acg aac ata aca ctt cag att     3552
Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile
   1170                1175                1180
gcc aca gat gaa gac agc gga atc ctc ctg tat aag ggt gac aaa gac     3600
Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp
1185               1190                1195                1200
cat atc gcg gta gaa ctc tat cgg ggg cgt gtt cgt gcc agc tat gac     3648
His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg Ala Ser Tyr Asp
               1205                1210                1215
acc ggc tct cat cca gct tct gcc att tac agt gtg gag aca atc aat     3696
Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn
           1220                1225                1230
gat gga aac ttc cac att gtg gaa cta ctt gcc ttg gat cag agt ctc     3744
Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu
       1235                1240                1245
tct ttg tcc gtg gat ggt ggg aac ccc aaa atc atc act aac ttg tca     3792
Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser
   1250                1255                1260
aag cag tcc act ctg aat ttt gac tct cca ctc tat gta gga ggc atg     3840
Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met
1265               1270                1275                1280
cca ggg aag agt aac gtg gca tct ctg cgc cag gcc cct ggg cag aac     3888
Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn
               1285                1290                1295
gga acc agc ttc cac ggc tgc atc cgg aac ctt tac atc aac agt gag     3936
Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu
           1300                1305                1310
ctg cag gac ttc cag aag gtg ccg atg caa aca ggc att ttg cct ggc     3984
Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly
       1315                1320                1325
tgt gag cca tgc cac aag aag gtg tgt gcc cat ggc aca tgc cag ccc     4032
Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro
   1330                1335                1340
agc agc cag gca ggc ttc acc tgc gag tgc cag gaa gga tgg atg ggg     4080
Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly
1345               1350                1355                1360
ccc ctc tgt gac caa cgg acc aat gac cct tgc ctt gga aat aaa tgc     4128
Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys
               1365                1370                1375
gta cat ggc acc tgc ttg ccc atc aat gcg ttc tcc tac agc tgt aag     4176
Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys
           1380                1385                1390
tgc ttg gag ggc cat gga ggt gtc ctc tgt gat gaa gag gag gat ctg     4224
Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu
       1395                1400                1405
ttt aac cca tgc cag gcg atc aag tgc aag cat ggg aag tgc agg ctt     4272
Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu
   1410                1415                1420
tca ggt ctg ggg cag ccc tac tgt gaa tgc agc agt gga tac acg ggg     4320
Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly
1425               1430                1435                1440
gac agc tgt gat cga gaa atc tct tgt cga ggg gaa agg ata aga gat     4368
Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp
               1445                1450                1455
tat tac caa aag cag cag ggc tat gct gct tgc caa aca acc aag aag     4416
Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys
           1460                1465                1470
gtg tcc cga tta gag tgc aga ggt ggg tgt gca gga ggg cag tgc tgt     4464
Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys
       1475                1480                1485
gga ccg ctg agg agc aag cgg cgg aaa tac tct ttc gaa tgc act gac     4512
Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp
   1490                1495                1500
ggc tcc tcc ttt gtg gac gag gtt gag aaa gtg gtg aag tgc ggc tgt     4560
Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys
1505               1510                1515                1520
acg agg tgt gtg tcc taaacacact cccggcagct ctgtctttgg aaaaggttgt     4615
Thr Arg Cys Val Ser
               1525
atacttcttg accatgtggg actaatgaat gcttcatagt ggaaatattt gaaatatatt   4675
gtaaaataca gaacagactt atttttatta tgagaataaa gacttttttt ctgcatttgg   4735
aaaaaaaaaa aaaaaaaact cga                                           4758 
           
             2 
             1525 
             PRT 
             human 
           
            2
Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val
  1               5                  10                  15
Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys
             20                  25                  30
Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser
         35                  40                  45
Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly
     50                  55                  60
Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His
 65                  70                  75                  80
Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg
                 85                  90                  95
Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg
            100                 105                 110
Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys
        115                 120                 125
Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg
    130                 135                 140
Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr
145                 150                 155                 160
Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp
                165                 170                 175
Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val
            180                 185                 190
Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser
        195                 200                 205
Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu
    210                 215                 220
Arg Lys Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser
225                 230                 235                 240
His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val
                245                 250                 255
Cys Ser Asp Glu Glu Glu Gly His Gln Ser Phe Met Ala Pro Ser Cys
            260                 265                 270
Ser Val Leu His Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val
        275                 280                 285
Asp Cys Arg Gly Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu
    290                 295                 300
Thr Ile Thr Glu Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro
305                 310                 315                 320
Pro Gly Ala Phe Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser
                325                 330                 335
Asn Asn Gln Ile Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg
            340                 345                 350
Ser Leu Asn Ser Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro
        355                 360                 365
Lys Ser Leu Phe Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn
    370                 375                 380
Ala Asn Lys Ile Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His
385                 390                 395                 400
Asn Leu Asn Leu Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala
                405                 410                 415
Lys Gly Thr Phe Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala
            420                 425                 430
Gln Asn Pro Phe Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr
        435                 440                 445
Leu His Thr Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro
    450                 455                 460
Arg Arg Leu Ala Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe
465                 470                 475                 480
Arg Cys Ser Gly Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys
                485                 490                 495
Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr
            500                 505                 510
Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro
        515                 520                 525
Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu
    530                 535                 540
Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn
545                 550                 555                 560
Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly
                565                 570                 575
Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn
            580                 585                 590
Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met
        595                 600                 605
Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly
    610                 615                 620
Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr
625                 630                 635                 640
Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn
                645                 650                 655
Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly
            660                 665                 670
Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln
        675                 680                 685
Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln
    690                 695                 700
Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu
705                 710                 715                 720
Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys
                725                 730                 735
Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val
            740                 745                 750
Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu
        755                 760                 765
Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg
    770                 775                 780
Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu
785                 790                 795                 800
Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr
                805                 810                 815
Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp
            820                 825                 830
Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser
        835                 840                 845
His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln
    850                 855                 860
Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala
865                 870                 875                 880
Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr
                885                 890                 895
Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu
            900                 905                 910
Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr
        915                 920                 925
Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly
    930                 935                 940
Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn
945                 950                 955                 960
Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp
                965                 970                 975
Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu
            980                 985                 990
Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr
        995                1000                1005
Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr
   1010                1015                1020
Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu
1025               1030                1035                1040
Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe
               1045                1050                1055
Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp
           1060                1065                1070
Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr
       1075                1080                1085
Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly
   1090                1095                1100
Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro
1105               1110                1115                1120
Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile
               1125                1130                1135
Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys
           1140                1145                1150
Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln
       1155                1160                1165
Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile
   1170                1175                1180
Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp
1185               1190                1195                1200
His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg Ala Ser Tyr Asp
               1205                1210                1215
Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn
           1220                1225                1230
Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu
       1235                1240                1245
Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser
   1250                1255                1260
Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met
1265               1270                1275                1280
Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn
               1285                1290                1295
Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu
           1300                1305                1310
Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly
       1315                1320                1325
Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro
   1330                1335                1340
Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly
1345               1350                1355                1360
Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys
               1365                1370                1375
Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys
           1380                1385                1390
Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu
       1395                1400                1405
Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu
   1410                1415                1420
Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly
1425               1430                1435                1440
Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp
               1445                1450                1455
Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys
           1460                1465                1470
Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys
       1475                1480                1485
Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp
   1490                1495                1500
Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys
1505               1510                1515                1520
Thr Arg Cys Val Ser
               1525 
           
             3 
             105 
             PRT 
             human 
           
            3
Ser Pro Cys Thr Cys Ser Asn Asn Ile Val Asp Cys Arg Gly Lys Gly
  1               5                  10                  15
Leu Met Glu Ile Pro Ala Asn Leu Pro Glu Gly Ile Val Glu Ile Arg
             20                  25                  30
Leu Glu Gln Asn Ser Ile Lys Ala Ile Pro Ala Gly Ala Phe Thr Gln
         35                  40                  45
Tyr Lys Lys Leu Lys Arg Ile Asp Ile Ser Lys Asn Gln Ile Ser Asp
     50                  55                  60
Ile Ala Pro Asp Ala Phe Gln Gly Leu Lys Ser Leu Thr Ser Leu Val
 65                  70                  75                  80
Leu Tyr Gly Asn Lys Ile Thr Glu Ile Ala Lys Gly Leu Phe Asp Gly
                 85                  90                  95
Leu Val Ser Leu Gln Leu Leu Leu Leu
            100                 105 
           
             4 
             138 
             PRT 
             human 
           
            4
Glu Gly Ala Phe Asn Gly Ala Ala Ser Val Gln Glu Leu Met Leu Thr
  1               5                  10                  15
Gly Asn Gln Leu Glu Thr Val His Gly Arg Gly Phe Arg Gly Gly Leu
             20                  25                  30
Ser Gly Leu Lys Thr Leu Met Leu Arg Ser Asn Leu Ile Gly Cys Val
         35                  40                  45
Ser Asn Asp Thr Phe Ala Gly Leu Ser Ser Val Arg Leu Leu Ser Leu
     50                  55                  60
Tyr Asp Asn Arg Ile Thr Thr Ile Thr Pro Gly Ala Phe Thr Thr Leu
 65                  70                  75                  80
Val Ser Leu Ser Thr Ile Asn Leu Leu Ser Asn Pro Phe Asn Cys Asn
                 85                  90                  95
Cys His Leu Gly Ala Gly Leu Gly Lys Trp Leu Arg Lys Arg Arg Ile
            100                 105                 110
Val Ser Gly Asn Pro Arg Cys Gln Lys Pro Phe Phe Leu Lys Glu Ile
        115                 120                 125
Pro Ile Gln Gly Val Gly His Pro Gly Ile
    130                 135 
           
             5 
             160 
             PRT 
             human 
             
               misc_feature 
               (121)..(150) 
               note=“Xaa signifies gap in sequence” 
             
           
            5
Trp Pro Arg Cys Glu Cys Met Pro Gly Tyr Ala Gly Asp Asn Cys Ser
  1               5                  10                  15
Glu Asn Gln Asp Asp Cys Arg Asp His Arg Cys Gln Asn Gly Ala Gln
             20                  25                  30
Cys Met Asp Glu Val Asn Ser Tyr Ser Cys Leu Cys Ala Glu Gly Tyr
         35                  40                  45
Ser Gly Gln Leu Cys Glu Ile Pro Pro His Leu Pro Ala Pro Lys Ser
     50                  55                  60
Pro Cys Glu Gly Thr Glu Cys Gln Asn Gly Ala Asn Cys Val Asp Gln
 65                  70                  75                  80
Gly Asn Arg Pro Val Cys Gln Cys Leu Pro Gly Phe Gly Gly Pro Glu
                 85                  90                  95
Cys Glu Lys Leu Leu Ser Val Asn Phe Val Asp Arg Asp Thr Tyr Leu
            100                 105                 110
Gln Phe Thr Asp Leu Gln Asn Trp Xaa Arg Xaa Asn Ile Thr Leu Gln
        115                 120                 125
Val Phe Thr Ala Glu Asp Asn Gly Ile Leu Leu Tyr Asn Gly Gly Asn
    130                 135                 140
Asp His Ile Ala Val Xaa Leu Tyr Xaa Gly His Val Arg Phe Ser Tyr
145                 150                 155                 160 
           
             6 
             103 
             PRT 
             human 
           
            6
Gln Cys His Ile Ser Asp Gln Gly Glu Pro Tyr Cys Leu Cys Gln Pro
  1               5                  10                  15
Gly Phe Ser Gly Glu His Cys Gln Gln Glu Asn Pro Cys Leu Gly Gln
             20                  25                  30
Val Val Arg Glu Val Ile Arg Arg Gln Lys Gly Tyr Ala Ser Cys Ala
         35                  40                  45
Thr Ala Ser Lys Val Pro Ile Met Glu Cys Arg Gly Gly Cys Gly Pro
     50                  55                  60
Gln Cys Cys Gln Pro Thr Arg Ser Lys Arg Arg Lys Tyr Val Phe Gln
 65                  70                  75                  80
Cys Thr Asp Gly Ser Ser Phe Val Glu Glu Val Glu Arg His Leu Glu
                 85                  90                  95
Cys Gly Cys Leu Ala Cys Ser
            100 
           
             7 
             1480 
             PRT 
             Drosophila melanogaster 
           
            7
Met Ala Ala Pro Ser Arg Thr Thr Leu Met Pro Pro Pro Phe Arg Leu
  1               5                  10                  15
Gln Leu Arg Leu Leu Ile Leu Pro Ile Leu Leu Leu Leu Arg His Asp
             20                  25                  30
Ala Val His Ala Glu Pro Tyr Ser Gly Gly Phe Gly Ser Ser Ala Val
         35                  40                  45
Ser Ser Gly Gly Leu Gly Ser Val Gly Ile His Ile Pro Gly Gly Gly
     50                  55                  60
Val Gly Val Ile Thr Glu Ala Arg Cys Pro Arg Val Cys Ser Cys Thr
 65                  70                  75                  80
Gly Leu Asn Val Asp Cys Ser His Arg Gly Leu Thr Ser Val Pro Arg
                 85                  90                  95
Lys Ile Ser Ala Asp Val Glu Arg Leu Glu Leu Gln Gly Asn Asn Leu
            100                 105                 110
Thr Val Ile Tyr Glu Thr Asp Phe Gln Arg Leu Thr Lys Leu Arg Met
        115                 120                 125
Leu Gln Leu Thr Asp Asn Gln Ile His Thr Ile Glu Arg Asn Ser Phe
    130                 135                 140
Gln Asp Leu Val Ser Leu Glu Arg Leu Asp Ile Ser Asn Asn Val Ile
145                 150                 155                 160
Thr Thr Val Gly Arg Arg Val Phe Lys Gly Ala Gln Ser Leu Arg Ser
                165                 170                 175
Leu Gln Leu Asp Asn Asn Gln Ile Thr Cys Leu Asp Glu His Ala Phe
            180                 185                 190
Lys Gly Leu Val Glu Leu Glu Ile Leu Thr Leu Asn Asn Asn Asn Leu
        195                 200                 205
Thr Ser Leu Pro His Asn Ile Phe Gly Gly Leu Gly Arg Leu Arg Ala
    210                 215                 220
Leu Arg Leu Ser Asp Asn Pro Phe Ala Cys Asp Cys His Leu Ser Trp
225                 230                 235                 240
Leu Ser Arg Phe Leu Arg Ser Ala Thr Arg Leu Ala Pro Tyr Thr Arg
                245                 250                 255
Cys Gln Ser Pro Ser Gln Leu Lys Gly Gln Asn Val Ala Asp Leu His
            260                 265                 270
Asp Gln Glu Phe Lys Cys Ser Gly Leu Thr Glu His Ala Pro Met Glu
        275                 280                 285
Cys Gly Ala Glu Asn Ser Cys Pro His Pro Cys Arg Cys Ala Asp Gly
    290                 295                 300
Ile Val Asp Cys Arg Glu Lys Ser Leu Thr Ser Val Pro Val Thr Leu
305                 310                 315                 320
Pro Asp Asp Thr Thr Asp Val Arg Leu Glu Gln Asn Phe Ile Thr Glu
                325                 330                 335
Leu Pro Pro Lys Ser Phe Ser Ser Phe Arg Arg Leu Arg Arg Ile Asp
            340                 345                 350
Leu Ser Asn Asn Asn Ile Ser Arg Ile Ala His Asp Ala Leu Ser Gly
        355                 360                 365
Leu Lys Gln Leu Thr Thr Leu Val Leu Tyr Gly Asn Lys Ile Lys Asp
    370                 375                 380
Leu Pro Ser Gly Val Phe Lys Gly Leu Gly Ser Leu Arg Leu Leu Leu
385                 390                 395                 400
Leu Asn Ala Asn Glu Ile Ser Cys Ile Arg Lys Asp Ala Phe Arg Asp
                405                 410                 415
Leu His Ser Leu Ser Leu Leu Ser Leu Tyr Asp Asn Asn Ile Gln Ser
            420                 425                 430
Leu Ala Asn Gly Thr Phe Asp Ala Met Lys Ser Met Lys Thr Val His
        435                 440                 445
Leu Ala Lys Asn Pro Phe Ile Cys Asp Cys Asn Leu Arg Trp Leu Ala
    450                 455                 460
Asp Tyr Leu His Lys Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Glu
465                 470                 475                 480
Ser Pro Lys Arg Met His Arg Arg Arg Ile Glu Ser Leu Arg Glu Glu
                485                 490                 495
Lys Phe Lys Cys Ser Trp Gly Glu Leu Arg Met Lys Leu Ser Gly Glu
            500                 505                 510
Cys Arg Met Asp Ser Asp Cys Pro Ala Met Cys His Cys Glu Gly Thr
        515                 520                 525
Thr Val Asp Cys Thr Gly Arg Arg Leu Lys Glu Ile Pro Arg Asp Ile
    530                 535                 540
Pro Leu His Thr Thr Glu Leu Leu Leu Asn Asp Asn Glu Leu Gly Arg
545                 550                 555                 560
Ile Ser Ser Asp Gly Leu Phe Gly Arg Leu Pro His Leu Val Lys Leu
                565                 570                 575
Glu Leu Lys Arg Asn Gln Leu Thr Gly Ile Glu Pro Asn Ala Phe Glu
            580                 585                 590
Gly Ala Ser His Ile Gln Glu Leu Gln Leu Gly Glu Asn Lys Ile Lys
        595                 600                 605
Glu Ile Ser Asn Lys Met Phe Leu Gly Leu His Gln Leu Lys Thr Leu
    610                 615                 620
Asn Leu Tyr Asp Asn Gln Ile Ser Cys Val Met Pro Gly Ser Phe Glu
625                 630                 635                 640
His Leu Asn Ser Leu Thr Ser Leu Asn Leu Ala Ser Asn Pro Phe Asn
                645                 650                 655
Cys Asn Cys His Leu Ala Trp Phe Ala Glu Cys Val Arg Lys Lys Ser
            660                 665                 670
Leu Asn Gly Gly Ala Ala Arg Cys Gly Ala Pro Ser Lys Val Arg Asp
        675                 680                 685
Val Gln Ile Lys Asp Leu Pro His Ser Glu Phe Lys Cys Ser Ser Glu
    690                 695                 700
Asn Ser Glu Gly Cys Leu Gly Asp Gly Tyr Cys Pro Pro Ser Cys Thr
705                 710                 715                 720
Cys Thr Gly Thr Val Val Ala Cys Ser Arg Asn Gln Leu Lys Glu Ile
                725                 730                 735
Pro Arg Gly Ile Pro Ala Glu Thr Ser Glu Leu Tyr Leu Glu Ser Asn
            740                 745                 750
Glu Ile Glu Gln Ile His Tyr Glu Arg Ile Arg His Leu Arg Ser Leu
        755                 760                 765
Thr Arg Leu Asp Leu Ser Asn Asn Gln Ile Thr Ile Leu Ser Asn Tyr
    770                 775                 780
Thr Phe Ala Asn Leu Thr Lys Leu Ser Thr Leu Ile Ile Ser Tyr Asn
785                 790                 795                 800
Lys Leu Gln Cys Leu Gln Arg His Ala Leu Ser Gly Leu Asn Asn Leu
                805                 810                 815
Arg Val Val Ser Leu His Gly Asn Arg Ile Ser Met Leu Pro Glu Gly
            820                 825                 830
Ser Phe Glu Asp Leu Lys Ser Leu Thr His Ile Ala Leu Gly Ser Asn
        835                 840                 845
Pro Leu Tyr Cys Asp Cys Gly Leu Lys Trp Phe Ser Asp Trp Ile Lys
    850                 855                 860
Leu Asp Tyr Val Glu Pro Gly Ile Ala Arg Cys Ala Glu Pro Glu Gln
865                 870                 875                 880
Met Lys Asp Lys Leu Ile Leu Ser Thr Pro Ser Ser Ser Phe Val Cys
                885                 890                 895
Arg Gly Arg Val Arg Asn Asp Ile Leu Ala Lys Cys Asn Ala Cys Phe
            900                 905                 910
Glu Gln Pro Cys Gln Asn Gln Ala Gln Cys Val Ala Leu Pro Gln Arg
        915                 920                 925
Glu Tyr Gln Cys Leu Cys Gln Pro Gly Tyr His Gly Lys His Cys Glu
    930                 935                 940
Phe Met Ile Asp Ala Cys Tyr Gly Asn Pro Cys Arg Asn Asn Ala Thr
945                 950                 955                 960
Cys Thr Val Leu Glu Glu Gly Arg Phe Ser Cys Gln Cys Ala Pro Gly
                965                 970                 975
Tyr Thr Gly Ala Arg Cys Glu Thr Asn Ile Asp Asp Cys Leu Gly Glu
            980                 985                 990
Ile Lys Cys Gln Asn Asn Ala Thr Cys Ile Asp Gly Val Glu Ser Tyr
        995                1000                1005
Lys Cys Glu Cys Gln Pro Gly Phe Ser Gly Glu Phe Cys Asp Thr Lys
   1010                1015                1020
Ile Gln Phe Cys Ser Pro Glu Phe Asn Pro Cys Ala Asn Gly Ala Lys
1025               1030                1035                1040
Cys Met Asp His Phe Thr His Tyr Ser Cys Asp Cys Gln Ala Gly Phe
               1045                1050                1055
His Gly Thr Asn Cys Thr Asp Asn Ile Asp Asp Cys Gln Asn His Met
           1060                1065                1070
Cys Gln Asn Gly Gly Thr Cys Val Asp Gly Ile Asn Asp Tyr Gln Cys
       1075                1080                1085
Arg Cys Pro Asp Asp Tyr Thr Gly Lys Tyr Cys Glu Gly His Asn Met
   1090                1095                1100
Ile Ser Met Met Tyr Pro Gln Thr Ser Pro Cys Gln Asn His Glu Cys
1105               1110                1115                1120
Lys His Gly Val Cys Phe Gln Pro Asn Ala Gln Gly Ser Asp Tyr Leu
               1125                1130                1135
Cys Arg Cys His Pro Gly Tyr Thr Gly Lys Trp Cys Glu Tyr Leu Thr
           1140                1145                1150
Ser Ile Ser Phe Val His Asn Asn Ser Phe Val Glu Leu Glu Pro Leu
       1155                1160                1165
Arg Thr Arg Pro Glu Ala Asn Val Thr Ile Val Phe Ser Ser Ala Glu
   1170                1175                1180
Gln Asn Gly Ile Leu Met Tyr Asp Gly Gln Asp Ala His Leu Ala Val
1185               1190                1195                1200
Glu Leu Phe Asn Gly Arg Ile Arg Val Ser Tyr Asp Val Gly Asn His
               1205                1210                1215
Pro Val Ser Thr Met Tyr Ser Phe Glu Met Val Ala Asp Gly Lys Tyr
           1220                1225                1230
His Ala Val Glu Leu Leu Ala Ile Lys Lys Asn Phe Thr Leu Arg Val
       1235                1240                1245
Asp Arg Gly Leu Ala Arg Ser Ile Ile Asn Glu Gly Ser Asn Asp Tyr
   1250                1255                1260
Leu Lys Leu Thr Thr Pro Met Phe Leu Gly Gly Leu Pro Val Asp Pro
1265               1270                1275                1280
Ala Gln Gln Ala Tyr Lys Asn Trp Gln Ile Arg Asn Leu Thr Ser Phe
               1285                1290                1295
Lys Gly Cys Met Lys Glu Val Trp Ile Asn His Lys Leu Val Asp Phe
           1300                1305                1310
Gly Asn Ala Gln Arg Gln Gln Lys Ile Thr Pro Gly Cys Ala Leu Leu
       1315                1320                1325
Glu Gly Glu Gln Gln Glu Glu Glu Asp Asp Glu Gln Asp Phe Met Asp
   1330                1335                1340
Glu Thr Pro His Ile Lys Glu Glu Pro Val Asp Pro Cys Leu Glu Asn
1345               1350                1355                1360
Lys Cys Arg Arg Gly Ser Arg Cys Val Pro Asn Ser Asn Ala Arg Asp
               1365                1370                1375
Gly Tyr Gln Cys Lys Cys Lys His Gly Gln Arg Gly Arg Tyr Cys Asp
           1380                1385                1390
Gln Gly Glu Gly Ser Thr Glu Pro Pro Thr Val Thr Ala Ala Ser Thr
       1395                1400                1405
Cys Arg Lys Glu Gln Val Arg Glu Tyr Tyr Thr Glu Asn Asp Cys Arg
   1410                1415                1420
Ser Arg Gln Pro Leu Lys Tyr Ala Lys Cys Val Gly Gly Cys Gly Asn
1425               1430                1435                1440
Gln Cys Cys Ala Ala Lys Ile Val Arg Arg Arg Lys Val Arg Met Val
               1445                1450                1455
Cys Ser Asn Asn Arg Lys Tyr Ile Lys Asn Leu Asp Ile Val Arg Lys
           1460                1465                1470
Cys Gly Cys Thr Lys Lys Cys Tyr
       1475                1480 
           
             8 
             155 
             PRT 
             Caenorhabditis elegans 
             
               misc_feature 
               (4)..(152) 
               note=“Xaa signifies gap in sequence” 
             
           
            8
Arg Asn Pro Xaa Ile Cys Asp Cys Asn Leu Gln Trp Leu Ala Gln Ile
  1               5                  10                  15
Asn Leu Gln Lys Asn Ile Glu Thr Ser Gly Ala Arg Cys Glu Gln Pro
             20                  25                  30
Lys Arg Leu Arg Lys Lys Lys Phe Ala Thr Leu Pro Pro Asn Lys Phe
         35                  40                  45
Lys Cys Lys Gly Ser Glu Ser Phe Val Ser Met Tyr Ala Asp Ser Cys
     50                  55                  60
Phe Ile Asp Ser Ile Cys Pro Thr Gln Cys Asp Cys Tyr Gly Thr Thr
 65                  70                  75                  80
Val Asp Cys Asn Lys Arg Gly Leu Asn Thr Ile Pro Thr Ser Ile Pro
                 85                  90                  95
Arg Phe Ala Thr Gln Leu Leu Leu Ser Gly Asn Asn Ile Ser Thr Val
            100                 105                 110
Asp Leu Asn Ser Asn Ile His Val Leu Glu Asn Leu Glu Xaa Leu Asp
        115                 120                 125
Leu Ser Asn Asn His Ile Thr Phe Ile Asn Asp Lys Ser Phe Glu Lys
    130                 135                 140
Leu Ser Lys Leu Arg Glu Leu Xaa Leu Asn Asp
145                 150                 155 
           
             9 
             735 
             PRT 
             Caenorhabditis elegans 
           
            9
Ser Asn Lys Asn Leu Thr Ser Phe Pro Ser Arg Ile Pro Phe Asp Thr
  1               5                  10                  15
Thr Glu Leu Tyr Leu Asp Ala Asn Tyr Ile Asn Glu Ile Pro Ala His
             20                  25                  30
Asp Leu Asn Arg Leu Tyr Ser Leu Thr Lys Leu Asp Leu Ser His Asn
         35                  40                  45
Arg Leu Ile Ser Leu Glu Asn Asn Thr Phe Ser Asn Leu Thr Arg Leu
     50                  55                  60
Ser Thr Leu Ile Ile Ser Tyr Asn Lys Leu Arg Cys Leu Gln Pro Leu
 65                  70                  75                  80
Ala Phe Asn Gly Leu Asn Ala Leu Arg Ile Leu Ser Leu His Gly Asn
                 85                  90                  95
Asp Ile Ser Phe Leu Pro Gln Ser Ala Phe Ser Asn Leu Thr Ser Ile
            100                 105                 110
Thr His Ile Ala Val Gly Ser Asn Ser Leu Tyr Cys Asp Cys Asn Met
        115                 120                 125
Ala Trp Phe Ser Lys Trp Ile Lys Ser Lys Phe Ile Glu Ala Gly Ile
    130                 135                 140
Ala Arg Cys Glu Tyr Pro Asn Thr Val Ser Asn Gln Leu Leu Leu Thr
145                 150                 155                 160
Ala Gln Pro Tyr Gln Phe Thr Cys Asp Ser Lys Val Pro Thr Lys Leu
                165                 170                 175
Ala Thr Lys Cys Asp Leu Cys Leu Asn Ser Pro Cys Lys Asn Asn Ala
            180                 185                 190
Ile Cys Glu Thr Thr Ser Ser Arg Lys Tyr Thr Cys Asn Cys Thr Pro
        195                 200                 205
Gly Phe Tyr Gly Val His Cys Glu Asn Gln Ile Asp Ala Cys Tyr Gly
    210                 215                 220
Ser Pro Cys Leu Asn Asn Ala Thr Cys Lys Val Ala Gln Ala Gly Arg
225                 230                 235                 240
Phe Asn Cys Tyr Cys Asn Lys Gly Phe Glu Gly Asp Tyr Cys Glu Lys
                245                 250                 255
Asn Ile Asp Asp Cys Val Asn Ser Lys Cys Glu Asn Gly Gly Lys Cys
            260                 265                 270
Val Asp Leu Val Arg Phe Cys Ser Glu Glu Leu Lys Asn Phe Gln Ser
        275                 280                 285
Phe Gln Ile Asn Ser Tyr Arg Cys Asp Cys Pro Met Glu Tyr Glu Gly
    290                 295                 300
Lys His Cys Glu Asp Lys Leu Glu Tyr Cys Thr Lys Lys Leu Asn Pro
305                 310                 315                 320
Cys Glu Asn Asn Gly Lys Cys Ile Pro Ile Asn Gly Ser Tyr Ser Cys
                325                 330                 335
Met Cys Ser Pro Gly Phe Thr Gly Asn Asn Cys Glu Thr Asn Ile Asp
            340                 345                 350
Asp Cys Lys Asn Val Glu Cys Gln Asn Gly Gly Ser Cys Val Asp Gly
        355                 360                 365
Ile Leu Ser Tyr Asp Cys Leu Cys Arg Pro Gly Tyr Ala Gly Gln Tyr
    370                 375                 380
Cys Glu Ile Pro Pro Met Met Asp Met Glu Tyr Gln Lys Thr Asp Ala
385                 390                 395                 400
Cys Gln Gln Ser Ala Cys Gly Gln Gly Glu Cys Val Ala Ser Gln Asn
                405                 410                 415
Ser Ser Asp Phe Thr Cys Lys Cys His Glu Gly Phe Ser Gly Pro Ser
            420                 425                 430
Cys Asp Arg Gln Met Ser Val Gly Phe Lys Asn Pro Gly Ala Tyr Leu
        435                 440                 445
Ala Leu Asp Pro Leu Ala Ser Asp Gly Thr Ile Thr Met Thr Leu Arg
    450                 455                 460
Thr Thr Ser Lys Ile Gly Ile Leu Leu Tyr Tyr Gly Asp Asp His Phe
465                 470                 475                 480
Val Ser Ala Glu Leu Tyr Asp Gly Arg Val Lys Leu Val Tyr Tyr Ile
                485                 490                 495
Gly Asn Phe Pro Ala Ser His Met Tyr Ser Ser Val Lys Val Asn Asp
            500                 505                 510
Gly Leu Pro His Arg Ile Ser Ile Arg Thr Ser Glu Arg Lys Cys Phe
        515                 520                 525
Leu Gln Ile Asp Lys Asn Pro Val Gln Ile Val Glu Asn Ser Gly Lys
    530                 535                 540
Ser Asp Gln Leu Ile Thr Lys Gly Lys Glu Met Leu Tyr Ile Gly Gly
545                 550                 555                 560
Leu Pro Ile Glu Lys Ser Gln Asp Ala Lys Arg Arg Phe His Val Lys
                565                 570                 575
Asn Ser Glu Ser Leu Lys Gly Cys Ile Ser Ser Ile Thr Ile Asn Glu
            580                 585                 590
Val Pro Ile Asn Leu Gln Gln Ala Leu Glu Asn Val Asn Thr Glu Gln
        595                 600                 605
Ser Cys Ser Ala Thr Val Asn Phe Cys Ala Gly Ile Asp Cys Gly Asn
    610                 615                 620
Gly Lys Cys Thr Asn Asn Ala Leu Ser Pro Lys Gly Tyr Met Cys Gln
625                 630                 635                 640
Cys Asp Ser His Phe Ser Gly Glu His Cys Asp Glu Lys Arg Ile Lys
                645                 650                 655
Cys Asp Lys Gln Lys Phe Arg Arg His His Ile Glu Asn Glu Cys Arg
            660                 665                 670
Ser Val Asp Arg Ile Lys Ile Ala Glu Cys Asn Gly Tyr Cys Gly Gly
        675                 680                 685
Glu Gln Asn Cys Cys Thr Ala Val Lys Lys Lys Gln Arg Lys Val Lys
    690                 695                 700
Met Ile Cys Lys Asn Gly Thr Thr Lys Ile Ser Thr Val His Ile Ile
705                 710                 715                 720
Arg Gln Cys Gln Cys Glu Pro Thr Lys Ser Val Leu Ser Glu Lys
                725                 730                 735 
           
             10 
             154 
             PRT 
             mouse 
           
            10
Asp Pro Leu Pro Val His His Arg Cys Glu Cys Met Leu Gly Tyr Thr
  1               5                  10                  15
Gly Asp Asn Cys Ser Glu Asn Gln Asp Asp Cys Lys Asp His Lys Cys
             20                  25                  30
Gln Asn Gly Ala Gln Cys Val Asp Glu Val Asn Ser Tyr Ala Cys Leu
         35                  40                  45
Cys Val Glu Gly Tyr Ser Gly Gln Leu Cys Glu Ile Pro Pro Ala Pro
     50                  55                  60
Arg Ser Ser Cys Glu Gly Thr Glu Cys Gln Asn Gly Ala Asn Cys Val
 65                  70                  75                  80
Asp Gln Gly Ser Arg Pro Val Cys Gln Cys Leu Pro Gly Phe Gly Gly
                 85                  90                  95
Pro Glu Cys Glu Lys Leu Leu Ser Val Asn Phe Val Asp Arg Asp Thr
            100                 105                 110
Tyr Leu Gln Phe Thr Asp Leu Gln Asn Trp Pro Arg Ala Asn Ile Thr
        115                 120                 125
Leu Gln Val Ser Thr Ala Glu Asp Asn Gly Ile Leu Leu Tyr Asn Gly
    130                 135                 140
Asp Asn Asp His Ile Ala Val Glu Leu Tyr
145                 150 
           
             11 
             110 
             PRT 
             mouse 
           
            11
Ala Phe Lys Cys His His Gly Gln Cys His Ile Ser Asp Arg Gly Glu
  1               5                  10                  15
Pro Tyr Cys Leu Cys Gln Pro Gly Phe Ser Gly His His Cys Glu Gln
             20                  25                  30
Glu Asn Pro Cys Met Gly Glu Ile Val Arg Glu Ala Ile Arg Arg Gln
         35                  40                  45
Lys Asp Tyr Ala Ser Cys Ala Thr Ala Ser Lys Val Pro Ile Met Glu
     50                  55                  60
Cys Arg Gly Gly Cys Gly Thr Thr Cys Cys Gln Pro Ile Arg Ser Lys
 65                  70                  75                  80
Arg Arg Lys Tyr Val Phe Gln Cys Thr Asp Gly Ser Ser Phe Val Glu
                 85                  90                  95
Glu Val Glu Arg His Leu Glu Cys Gly Cys Arg Ala Cys Ser
            100                 105                 110 
           
             12 
             134 
             PRT 
             mouse 
           
            12
His Leu Arg Val Leu Gln Leu Met Glu Asn Arg Ile Ser Thr Ile Glu
  1               5                  10                  15
Arg Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn
             20                  25                  30
Arg Asn Asn Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala
         35                  40                  45
Arg Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro
     50                  55                  60
Arg Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp
 65                  70                  75                  80
Tyr Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg
                 85                  90                  95
Asp Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser
            100                 105                 110
Val Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His
        115                 120                 125
Ser Asn Asn Leu Tyr Cys
    130 
           
             13 
             104 
             PRT 
             mouse 
           
            13
Asn Asn Asp Asp Cys Val Gly His Lys Cys Arg His Gly Ala Gln Cys
  1               5                  10                  15
Val Asp Glu Val Asn Gly Tyr Thr Cys Ile Cys Pro Gln Gly Phe Ser
             20                  25                  30
Gly Leu Phe Cys Glu His Pro Pro Pro Met Val Leu Leu Gln Thr Ser
         35                  40                  45
Pro Cys Asp Gln Tyr Glu Cys Gln Asn Gly Ala Gln Cys Ile Val Val
     50                  55                  60
Gln Gln Glu Pro Thr Cys Arg Cys Pro Pro Gly Phe Ala Gly Pro Arg
 65                  70                  75                  80
Cys Glu Lys Leu Ile Thr Val Asn Phe Val Gly Lys Asp Ser Tyr Val
                 85                  90                  95
Glu Leu Ala Ser Ala Lys Val Arg
            100 
           
             14 
             243 
             PRT 
             mouse 
           
            14
Ile Leu Asp Val Ala Ser Leu Arg Gln Ala Pro Gly Glu Asn Gly Thr
  1               5                  10                  15
Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu Leu Gln
             20                  25                  30
Asp Phe Arg Lys Met Pro Met Gln Thr Gly Ile Leu Pro Gly Cys Glu
         35                  40                  45
Pro Cys His Lys Lys Val Cys Ala His Gly Cys Cys Gln Pro Ser Ser
     50                  55                  60
Gln Ser Gly Phe Thr Cys Glu Cys Glu Glu Gly Trp Met Gly Pro Leu
 65                  70                  75                  80
Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys Val His
                 85                  90                  95
Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys Cys Leu
            100                 105                 110
Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu Phe Asn
        115                 120                 125
Pro Cys Gln Met Ile Lys Cys Lys His Gly Lys Cys Arg Leu Ser Gly
    130                 135                 140
Val Gly Gln Pro Tyr Cys Glu Cys Asn Ser Gly Phe Thr Gly Asp Ser
145                 150                 155                 160
Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp Tyr Tyr
                165                 170                 175
Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys Val Ser
            180                 185                 190
Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys Gly Pro
        195                 200                 205
Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp Gly Ser
    210                 215                 220
Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys Ala Arg
225                 230                 235                 240
Cys Ala Ser 
           
             15 
             1395 
             PRT 
             Drosophila melanogaster 
           
            15
Met His Pro Met His Pro Glu Asn His Ala Ile Ala Arg Ser Thr Ser
1               5                   10                  15
Thr Thr Asn Asn Pro Ser Arg Ser Arg Ser Ser Arg Met Trp Leu Leu
            20                  25                  30
Pro Ala Trp Leu Leu Leu Val Leu Val Ala Ser Asn Gly Leu Pro Ala
        35                  40                  45
Val Arg Gly Gln Tyr Gln Ser Pro Arg Ile Ile Glu His Pro Thr Asp
    50                  55                  60
Leu Val Val Lys Lys Asn Glu Pro Ala Thr Leu Asn Cys Lys Val Glu
65                  70                  75                  80
Gly Lys Pro Glu Pro Thr Ile Glu Trp Phe Lys Asp Gly Glu Pro Val
                85                  90                  95
Ser Thr Asn Glu Lys Lys Ser His Arg Val Gln Phe Lys Asp Gly Ala
            100                 105                 110
Leu Phe Phe Tyr Arg Thr Met Gln Gly Lys Lys Glu Gln Asp Gly Gly
        115                 120                 125
Glu Tyr Trp Cys Val Ala Lys Asn Arg Val Gly Gln Ala Val Ser Arg
    130                 135                 140
His Ala Ser Leu Gln Ile Ala Val Leu Arg Asp Asp Phe Arg Val Glu
145                 150                 155                 160
Pro Lys Asp Thr Arg Val Ala Lys Gly Glu Thr Ala Leu Leu Glu Cys
                165                 170                 175
Gly Pro Pro Lys Gly Ile Pro Glu Pro Thr Leu Ile Trp Ile Lys Asp
            180                 185                 190
Gly Val Pro Leu Asp Asp Leu Lys Ala Met Ser Phe Gly Ala Ser Ser
        195                 200                 205
Arg Val Arg Ile Val Asp Gly Gly Asn Leu Leu Ile Ser Asn Val Glu
    210                 215                 220
Pro Ile Asp Glu Gly Asn Tyr Lys Cys Ile Ala Gln Asn Leu Val Gly
225                 230                 235                 240
Thr Arg Glu Ser Ser Tyr Ala Lys Leu Ile Val Gln Val Lys Pro Tyr
                245                 250                 255
Phe Met Lys Glu Pro Lys Asp Gln Val Met Leu Tyr Gly Gln Thr Ala
            260                 265                 270
Thr Phe His Cys Ser Val Gly Gly Asp Pro Pro Pro Lys Val Leu Trp
        275                 280                 285
Lys Lys Glu Glu Gly Asn Ile Pro Val Ser Arg Ala Arg Ile Leu His
    290                 295                 300
Asp Glu Lys Ser Leu Glu Ile Ser Asn Ile Thr Pro Thr Asp Glu Gly
305                 310                 315                 320
Thr Tyr Val Cys Glu Ala His Asn Asn Val Gly Gln Ile Ser Ala Arg
                325                 330                 335
Ala Ser Leu Ile Val His Ala Pro Pro Asn Phe Thr Lys Arg Pro Ser
            340                 345                 350
Asn Lys Lys Val Gly Leu Asn Gly Val Val Gln Leu Pro Cys Met Ala
        355                 360                 365
Ser Gly Asn Pro Pro Pro Ser Val Phe Trp Thr Lys Glu Gly Val Ser
    370                 375                 380
Thr Leu Met Phe Pro Asn Ser Ser His Gly Arg Gln Tyr Val Ala Ala
385                 390                 395                 400
Asp Gly Thr Leu Gln Ile Thr Asp Val Arg Gln Glu Asp Glu Gly Tyr
                405                 410                 415
Tyr Val Cys Ser Ala Phe Ser Val Val Asp Ser Ser Thr Val Arg Val
            420                 425                 430
Phe Leu Gln Val Ser Ser Val Asp Glu Arg Pro Pro Pro Ile Ile Gln
        435                 440                 445
Ile Gly Pro Ala Asn Gln Thr Leu Pro Lys Gly Ser Val Ala Thr Leu
    450                 455                 460
Pro Cys Arg Ala Thr Gly Asn Pro Ser Pro Arg Ile Lys Trp Phe His
465                 470                 475                 480
Asp Gly His Ala Val Gln Ala Gly Asn Arg Tyr Ser Ile Ile Gln Gly
                485                 490                 495
Ser Ser Leu Arg Val Asp Asp Leu Gln Leu Ser Asp Ser Gly Thr Tyr
            500                 505                 510
Thr Cys Thr Ala Ser Gly Glu Arg Gly Glu Thr Ser Trp Ala Ala Thr
        515                 520                 525
Leu Thr Val Glu Lys Pro Gly Ser Thr Ser Leu His Arg Ala Ala Asp
    530                 535                 540
Pro Ser Thr Tyr Pro Ala Pro Pro Gly Thr Pro Lys Val Leu Asn Val
545                 550                 555                 560
Ser Arg Thr Ser Ile Ser Leu Arg Trp Ala Lys Ser Gln Glu Lys Pro
                565                 570                 575
Gly Ala Val Gly Pro Ile Ile Gly Tyr Thr Val Glu Tyr Phe Ser Pro
            580                 585                 590
Asp Leu Gln Thr Gly Trp Ile Val Ala Ala His Arg Val Gly Asp Thr
        595                 600                 605
Gln Val Thr Ile Ser Gly Leu Thr Pro Gly Thr Ser Tyr Val Phe Leu
    610                 615                 620
Val Arg Ala Glu Asn Thr Gln Gly Ile Ser Val Pro Ser Gly Leu Ser
625                 630                 635                 640
Asn Val Ile Lys Thr Ile Glu Ala Asp Phe Asp Ala Ala Ser Ala Asn
                645                 650                 655
Asp Leu Ser Ala Ala Arg Thr Leu Leu Thr Gly Lys Ser Val Glu Leu
            660                 665                 670
Ile Asp Ala Ser Ala Ile Asn Ala Ser Ala Val Arg Leu Glu Trp Met
        675                 680                 685
Leu His Val Ser Ala Asp Glu Lys Tyr Val Glu Gly Leu Arg Ile His
    690                 695                 700
Tyr Lys Asp Ala Ser Val Pro Ser Ala Gln Tyr His Ser Ile Thr Val
705                 710                 715                 720
Met Asp Ala Ser Ala Glu Ser Phe Val Val Gly Asn Leu Lys Lys Tyr
                725                 730                 735
Thr Lys Tyr Glu Phe Phe Leu Thr Pro Phe Phe Glu Thr Ile Glu Gly
            740                 745                 750
Gln Pro Ser Asn Ser Lys Thr Ala Leu Thr Tyr Glu Asp Val Pro Ser
        755                 760                 765
Ala Pro Pro Asp Asn Ile Gln Ile Gly Met Tyr Asn Gln Thr Ala Gly
    770                 775                 780
Trp Val Arg Trp Thr Pro Pro Pro Ser Gln His His Asn Gly Asn Leu
785                 790                 795                 800
Tyr Gly Tyr Lys Ile Glu Val Ser Ala Gly Asn Thr Met Lys Val Leu
                805                 810                 815
Ala Asn Met Thr Leu Asn Ala Thr Thr Thr Ser Val Leu Leu Asn Asn
            820                 825                 830
Leu Thr Thr Gly Ala Val Tyr Ser Val Arg Leu Asn Ser Phe Thr Lys
        835                 840                 845
Ala Gly Asp Gly Pro Tyr Ser Lys Pro Ile Ser Leu Phe Met Asp Pro
    850                 855                 860
Thr His His Val His Pro Pro Arg Ala His Pro Ser Gly Thr His Asp
865                 870                 875                 880
Gly Arg His Glu Gly Gln Asp Leu Thr Tyr His Asn Asn Gly Asn Ile
                885                 890                 895
Pro Pro Gly Asp Ile Asn Pro Thr Thr His Lys Lys Thr Thr Asp Tyr
            900                 905                 910
Leu Ser Gly Pro Trp Leu Met Val Leu Val Cys Ile Val Leu Leu Val
        915                 920                 925
Leu Val Ile Ser Ala Ala Ile Ser Met Val Tyr Phe Lys Arg Lys His
    930                 935                 940
Gln Met Thr Lys Glu Leu Gly His Leu Ser Val Val Ser Asp Asn Glu
945                 950                 955                 960
Ile Thr Ala Leu Asn Ile Asn Ser Lys Glu Ser Leu Trp Ile Asp His
                965                 970                 975
His Arg Gly Trp Arg Thr Ala Asp Thr Asp Lys Asp Ser Gly Leu Ser
            980                 985                 990
Glu Ser Lys Leu Leu Ser His Val Asn Ser Ser Gln Ser Asn Tyr Asn
        995                 1000                1005
Asn Ser Asp Gly Gly Thr Asp Tyr Ala Glu Val Asp Thr Arg Asn Leu
    1010                1015                1020
Thr Thr Phe Tyr Asn Cys Arg Lys Ser Pro Asp Asn Pro Thr Pro Tyr
1025                1030                1035                1040
Ala Thr Thr Met Ile Ile Gly Thr Ser Ser Ser Glu Thr Cys Thr Lys
                1045                1050                1055
Thr Thr Ser Ile Ser Ala Asp Lys Asp Ser Gly Thr His Ser Pro Tyr
            1060                1065                1070
Ser Asp Ala Phe Ala Gly Gln Val Pro Ala Val Pro Val Val Lys Ser
        1075                1080                1085
Asn Tyr Leu Gln Tyr Pro Val Glu Pro Ile Asn Trp Ser Glu Phe Leu
    1090                1095                1100
Pro Pro Pro Pro Glu His Pro Pro Pro Ser Ser Thr Tyr Gly Tyr Ala
1105                1110                1115                1120
Gln Gly Ser Pro Glu Ser Ser Arg Lys Ser Ser Lys Ser Ala Gly Ser
                1125                1130                1135
Gly Ile Ser Thr Asn Gln Ser Ile Leu Asn Ala Ser Ile His Ser Ser
            1140                1145                1150
Ser Ser Gly Gly Phe Ser Ala Trp Gly Val Ser Pro Gln Tyr Ala Val
        1155                1160                1165
Ala Cys Pro Pro Glu Asn Val Tyr Ser Asn Pro Leu Ser Ala Val Ala
    1170                1175                1180
Gly Gly Thr Gln Asn Arg Tyr Gln Ile Thr Pro Thr Asn Gln His Pro
1185                1190                1195                1200
Pro Gln Leu Pro Ala Tyr Phe Ala Thr Thr Gly Pro Gly Gly Ala Val
                1205                1210                1215
Pro Pro Asn His Leu Pro Phe Ala Thr Gln Arg His Ala Ala Ser Glu
            1220                1225                1230
Tyr Gln Ala Gly Leu Asn Ala Ala Arg Cys Ala Gln Ser Arg Ala Cys
        1235                1240                1245
Asn Ser Cys Asp Ala Leu Ala Thr Pro Ser Pro Met Gln Pro Pro Pro
    1250                1255                1260
Pro Val Pro Val Pro Glu Gly Trp Tyr Gln Pro Val His Pro Asn Ser
1265                1270                1275                1280
His Pro Met His Pro Thr Ser Ser Asn His Gln Ile Tyr Gln Cys Ser
                1285                1290                1295
Ser Glu Cys Ser Asp His Ser Arg Ser Ser Gln Ser His Lys Arg Gln
            1300                1305                1310
Leu Gln Leu Glu Glu His Gly Ser Ser Ala Lys Gln Arg Gly Gly His
        1315                1320                1325
His Arg Arg Arg Ala Pro Val Val Gln Pro Cys Met Glu Ser Glu Asn
    1330                1335                1340
Glu Asn Met Leu Ala Glu Tyr Glu Gln Arg Gln Tyr Thr Ser Asp Cys
1345                1350                1355                1360
Cys Asn Ser Ser Arg Glu Gly Asp Thr Cys Ser Cys Ser Glu Gly Ser
                1365                1370                1375
Cys Leu Tyr Ala Glu Ala Gly Glu Pro Ala Pro Arg Gln Met Thr Ala
            1380                1385                1390
Lys Asn Thr
        1395 
           
             16 
             1381 
             PRT 
             Drosophila melanogaster 
           
            16
Gly Glu Asn Pro Arg Ile Ile Glu His Pro Met Asp Thr Thr Val Pro
1               5                   10                  15
Lys Asn Asp Pro Phe Thr Phe Asn Cys Gln Ala Glu Gly Asn Pro Thr
            20                  25                  30
Pro Thr Ile Gln Trp Phe Lys Asp Gly Arg Glu Leu Lys Thr Asp Thr
        35                  40                  45
Gly Ser His Arg Ile Met Leu Pro Ala Gly Gly Leu Phe Phe Leu Lys
    50                  55                  60
Val Ile His Ser Arg Arg Glu Ser Asp Ala Gly Thr Tyr Trp Cys Glu
65                  70                  75                  80
Ala Lys Asn Glu Phe Gly Val Ala Arg Ser Arg Asn Ala Thr Leu Gln
                85                  90                  95
Val Ala Val Leu Arg Asp Glu Phe Arg Leu Glu Pro Ala Asn Thr Arg
            100                 105                 110
Val Ala Gln Gly Glu Val Ala Leu Met Glu Cys Gly Ala Pro Arg Gly
        115                 120                 125
Ser Pro Glu Pro Gln Ile Ser Trp Arg Lys Asn Gly Gln Thr Leu Asn
    130                 135                 140
Leu Val Gly Asn Lys Arg Ile Arg Ile Val Asp Gly Gly Asn Leu Ala
145                 150                 155                 160
Ile Gln Glu Ala Arg Gln Ser Asp Asp Gly Arg Tyr Gln Cys Val Val
                165                 170                 175
Lys Asn Val Val Gly Thr Arg Glu Ser Ala Thr Ala Phe Leu Lys Val
            180                 185                 190
His Val Arg Pro Phe Leu Ile Arg Gly Pro Gln Asn Gln Thr Ala Val
        195                 200                 205
Val Gly Ser Ser Val Val Phe Gln Cys Arg Ile Gly Gly Asp Pro Leu
    210                 215                 220
Pro Asp Val Leu Trp Arg Arg Thr Ala Ser Gly Gly Asn Met Pro Leu
225                 230                 235                 240
Arg Lys Phe Ser Trp Leu His Ser Ala Ser Gly Arg Val His Val Leu
                245                 250                 255
Glu Asp Arg Ser Leu Lys Leu Asp Asp Val Thr Leu Glu Asp Met Gly
            260                 265                 270
Glu Tyr Thr Cys Glu Ala Asp Asn Ala Val Gly Gly Ile Thr Ala Thr
        275                 280                 285
Gly Ile Leu Thr Val His Ala Pro Pro Lys Phe Val Ile Arg Pro Lys
    290                 295                 300
Asn Gln Leu Val Glu Ile Gly Asp Glu Val Leu Phe Glu Cys Gln Ala
305                 310                 315                 320
Asn Gly His Pro Arg Pro Thr Leu Tyr Trp Ser Val Glu Gly Asn Ser
                325                 330                 335
Ser Leu Leu Leu Pro Gly Tyr Arg Asp Gly Arg Met Glu Val Thr Leu
            340                 345                 350
Thr Pro Glu Gly Arg Ser Val Leu Ser Ile Ala Arg Phe Ala Arg Glu
        355                 360                 365
Asp Ser Gly Lys Val Val Thr Cys Asn Ala Leu Asn Ala Val Gly Ser
    370                 375                 380
Val Ser Ser Arg Thr Val Val Ser Val Asp Thr Gln Phe Glu Leu Pro
385                 390                 395                 400
Pro Pro Ile Ile Glu Gln Gly Pro Val Asn Gln Thr Leu Pro Val Lys
                405                 410                 415
Ser Ile Val Val Leu Pro Cys Arg Thr Leu Gly Thr Pro Val Pro Gln
            420                 425                 430
Val Ser Trp Tyr Leu Asp Gly Ile Pro Ile Asp Val Gln Glu His Glu
        435                 440                 445
Arg Arg Asn Leu Ser Asp Ala Gly Ala Leu Thr Ile Ser Asp Leu Gln
    450                 455                 460
Arg His Glu Asp Glu Gly Leu Tyr Thr Cys Val Ala Ser Asn Arg Asn
465                 470                 475                 480
Gly Lys Ser Ser Trp Ser Gly Tyr Leu Arg Leu Asp Thr Pro Thr Asn
                485                 490                 495
Pro Asn Ile Lys Phe Phe Arg Ala Pro Glu Leu Ser Thr Tyr Pro Gly
            500                 505                 510
Pro Pro Gly Lys Pro Gln Met Val Glu Lys Gly Glu Asn Ser Val Thr
        515                 520                 525
Leu Ser Trp Thr Arg Ser Asn Lys Val Gly Gly Ser Ser Leu Val Gly
    530                 535                 540
Tyr Val Ile Glu Met Phe Gly Lys Asn Glu Thr Asp Gly Trp Val Ala
545                 550                 555                 560
Val Gly Thr Arg Val Gln Asn Thr Thr Phe Thr Gln Thr Gly Leu Leu
                565                 570                 575
Pro Gly Val Asn Tyr Phe Phe Leu Ile Arg Ala Glu Asn Ser His Gly
            580                 585                 590
Leu Ser Leu Pro Ser Pro Met Ser Glu Pro Ile Thr Val Gly Thr Arg
        595                 600                 605
Tyr Phe Asn Ser Gly Leu Asp Leu Ser Glu Ala Arg Ala Ser Leu Leu
    610                 615                 620
Ser Gly Asp Val Val Glu Leu Ser Asn Ala Ser Val Val Asp Ser Thr
625                 630                 635                 640
Ser Met Lys Leu Thr Trp Gln Ile Ile Asn Gly Lys Tyr Val Glu Gly
                645                 650                 655
Phe Tyr Val Tyr Ala Arg Gln Leu Pro Asn Pro Ile Val Asn Asn Pro
            660                 665                 670
Ala Pro Val Thr Ser Asn Thr Asn Pro Leu Leu Gly Ser Thr Ser Thr
        675                 680                 685
Ser Ala Ser Ala Ser Ala Ser Ala Ser Ala Leu Ile Ser Thr Lys Pro
    690                 695                 700
Asn Ile Ala Ala Ala Gly Lys Arg Asp Gly Glu Thr Asn Gln Ser Gly
705                 710                 715                 720
Gly Gly Ala Pro Thr Pro Leu Asn Thr Lys Tyr Arg Met Leu Thr Ile
                725                 730                 735
Leu Asn Gly Gly Gly Ala Ser Ser Cys Thr Ile Thr Gly Leu Val Gln
            740                 745                 750
Tyr Thr Leu Tyr Glu Phe Phe Ile Val Pro Phe Tyr Lys Ser Val Glu
        755                 760                 765
Gly Lys Pro Ser Asn Ser Arg Ile Ala Arg Thr Leu Glu Asp Val Pro
    770                 775                 780
Ser Glu Ala Pro Tyr Gly Met Glu Ala Leu Leu Leu Asn Ser Ser Ala
785                 790                 795                 800
Val Phe Leu Lys Trp Lys Ala Pro Glu Leu Lys Asp Arg His Gly Val
                805                 810                 815
Leu Leu Asn Tyr His Val Ile Val Arg Gly Ile Asp Thr Ala His Asn
            820                 825                 830
Phe Ser Arg Ile Leu Thr Asn Val Thr Ile Asp Ala Ala Ser Pro Thr
        835                 840                 845
Leu Val Leu Ala Asn Leu Thr Glu Gly Val Met Tyr Thr Val Gly Val
    850                 855                 860
Ala Ala Gly Asn Asn Ala Gly Val Gly Pro Tyr Cys Val Pro Ala Thr
865                 870                 875                 880
Leu Arg Leu Asp Pro Ile Thr Lys Arg Leu Asp Pro Phe Ile Asn Gln
                885                 890                 895
Arg Asp His Val Asn Asp Val Leu Thr Gln Pro Trp Phe Ile Ile Leu
            900                 905                 910
Leu Gly Ala Ile Leu Ala Val Leu Met Leu Ser Phe Gly Ala Met Val
        915                 920                 925
Phe Val Lys Arg Lys His Met Met Met Lys Gln Ser Ala Leu Asn Thr
    930                 935                 940
Met Arg Gly Asn His Thr Ser Asp Val Leu Lys Met Pro Ser Leu Ser
945                 950                 955                 960
Ala Arg Asn Gly Asn Gly Tyr Trp Leu Asp Ser Ser Thr Gly Gly Met
                965                 970                 975
Val Trp Arg Pro Ser Pro Gly Gly Asp Ser Leu Glu Met Gln Lys Asp
            980                 985                 990
His Ile Ala Asp Tyr Ala Pro Val Cys Gly Ala Pro Gly Ser Pro Ala
        995                 1000                1005
Gly Gly Gly Thr Ser Ser Gly Gly Ser Gly Gly Ala Gly Ser Gly Ala
    1010                1015                1020
Ser Gly Gly Asp Asp Ile His Gly Gly His Gly Ser Glu Arg Asn Gln
1025                1030                1035                1040
Gln Arg Tyr Val Gly Glu Tyr Ser Asn Ile Pro Thr Asp Tyr Ala Glu
                1045                1050                1055
Val Ser Ser Phe Gly Lys Ala Pro Ser Glu Tyr Gly Arg His Gly Asn
            1060                1065                1070
Ala Ser Pro Ala Pro Tyr Ala Thr Ser Ser Ile Leu Ser Pro His Gln
        1075                1080                1085
Gln Gln Gln Gln Gln Gln Pro Arg Tyr Gln Gln Arg Pro Val Pro Gly
    1090                1095                1100
Tyr Gly Leu Gln Arg Pro Met His Pro His Tyr Gln Gln Gln Gln His
1105                1110                1115                1120
Gln Gln Gln Gln Ala Gln Gln Thr His Gln Gln His Gln Ala Leu Gln
                1125                1130                1135
Gln His Gln Gln Leu Pro Pro Ser Asn Ile Tyr Gln Gln Met Ser Thr
            1140                1145                1150
Thr Ser Glu Ile Tyr Pro Thr Asn Thr Gly Pro Ser Arg Ser Val Tyr
        1155                1160                1165
Ser Glu Gln Tyr Tyr Tyr Pro Lys Asp Lys Gln Arg His Ile His Ile
    1170                1175                1180
Thr Glu Asn Lys Leu Ser Asn Cys His Thr Tyr Glu Ala Ala Pro Gly
1185                1190                1195                1200
Ala Lys Gln Ser Ser Pro Ile Ser Ser Gln Phe Ala Ser Val Arg Arg
                1205                1210                1215
Gln Gln Leu Pro Pro Asn Cys Ser Ile Gly Arg Glu Ser Ala Arg Phe
            1220                1225                1230
Lys Val Leu Asn Thr Asp Gln Gly Lys Asn Gln Gln Asn Leu Leu Asp
        1235                1240                1245
Leu Asp Gly Ser Ser Met Cys Tyr Asn Gly Leu Ala Asp Ser Gly Cys
    1250                1255                1260
Gly Gly Ser Pro Ser Pro Met Ala Met Leu Met Ser His Glu Asp Glu
1265                1270                1275                1280
His Ala Leu Tyr His Thr Ala Asp Gly Asp Leu Asp Asp Met Glu Arg
                1285                1290                1295
Leu Tyr Val Lys Val Asp Glu Gln Gln Pro Pro Gln Gln Gln Gln Gln
            1300                1305                1310
Leu Ile Pro Leu Val Pro Gln His Pro Ala Glu Gly His Leu Gln Ser
        1315                1320                1325
Trp Arg Asn Gln Ser Thr Arg Ser Ser Arg Lys Asn Gly Gln Glu Cys
    1330                1335                1340
Ile Lys Glu Pro Ser Glu Leu Ile Tyr Ala Pro Gly Ser Val Ala Ser
1345                1350                1355                1360
Glu Arg Ser Leu Leu Ser Asn Ser Gly Ser Gly Thr Ser Ser Gln Pro
                1365                1370                1375
Ala Gly His Asn Val
            1380 
           
             17 
             1297 
             PRT 
             Caenorhabditis elegans 
           
            17
Met Tyr Tyr Leu Gly Phe Tyr His Thr His Thr His Thr His Thr Tyr
1               5                   10                  15
Ile Asn Phe Asp Lys Ile Pro Asn Ala Ser Asn Leu Ala Pro Val Ile
            20                  25                  30
Ile Glu His Pro Ile Asp Val Val Val Ser Arg Gly Ser Pro Ala Thr
        35                  40                  45
Leu Asn Cys Gly Ala Lys Pro Ser Thr Ala Lys Ile Thr Trp Tyr Lys
    50                  55                  60
Asp Gly Gln Pro Val Ile Thr Asn Lys Glu Gln Val Asn Ser His Arg
65                  70                  75                  80
Ile Val Leu Asp Thr Gly Ser Leu Phe Leu Leu Lys Val Asn Ser Gly
                85                  90                  95
Lys Asn Gly Lys Asp Ser Asp Ala Gly Ala Tyr Tyr Cys Val Ala Ser
            100                 105                 110
Asn Glu His Gly Glu Val Lys Ser Asn Glu Gly Ser Leu Lys Leu Ala
        115                 120                 125
Met Leu Arg Glu Asp Phe Arg Val Arg Pro Arg Thr Val Gln Ala Leu
    130                 135                 140
Gly Gly Glu Met Ala Val Leu Glu Cys Ser Pro Pro Arg Gly Phe Pro
145                 150                 155                 160
Glu Pro Val Val Ser Trp Arg Lys Asp Asp Lys Glu Leu Arg Ile Gln
                165                 170                 175
Asp Met Pro Arg Tyr Thr Leu His Ser Asp Gly Asn Leu Ile Ile Asp
            180                 185                 190
Pro Val Asp Arg Ser Asp Ser Gly Thr Tyr Gln Cys Val Ala Asn Asn
        195                 200                 205
Met Val Gly Glu Arg Val Ser Asn Pro Ala Arg Leu Ser Val Phe Glu
    210                 215                 220
Lys Pro Lys Phe Glu Gln Glu Pro Lys Asp Met Thr Val Asp Val Gly
225                 230                 235                 240
Ala Ala Val Leu Phe Asp Cys Arg Val Thr Gly Asp Pro Gln Pro Gln
                245                 250                 255
Ile Thr Trp Lys Arg Lys Asn Glu Pro Met Pro Val Thr Arg Ala Tyr
            260                 265                 270
Ile Ala Lys Asp Asn Arg Gly Leu Arg Ile Glu Arg Val Gln Pro Ser
        275                 280                 285
Asp Glu Gly Glu Tyr Val Cys Tyr Ala Arg Asn Pro Ala Gly Thr Leu
    290                 295                 300
Glu Ala Ser Ala His Leu Arg Val Gln Ala Pro Pro Ser Phe Gln Thr
305                 310                 315                 320
Lys Pro Ala Asp Gln Ser Val Pro Ala Gly Gly Thr Ala Thr Phe Glu
                325                 330                 335
Cys Thr Leu Val Gly Gln Pro Ser Pro Ala Tyr Phe Trp Ser Lys Glu
            340                 345                 350
Gly Gln Gln Asp Leu Leu Phe Pro Ser Tyr Val Ser Ala Asp Gly Arg
        355                 360                 365
Thr Lys Val Ser Pro Thr Gly Thr Leu Thr Ile Glu Glu Val Arg Gln
    370                 375                 380
Val Asp Glu Gly Ala Tyr Val Cys Ala Gly Met Asn Ser Ala Gly Ser
385                 390                 395                 400
Ser Leu Ser Lys Ala Ala Leu Lys Ala Thr Phe Glu Thr Lys Gly Arg
                405                 410                 415
Val Gln Lys Lys Lys Ser Lys Met Gly Lys Gln Lys Gln Lys Asn Val
            420                 425                 430
Gln Ser Ile Ile Lys Tyr Leu Ile Ser Ala Val Thr Gly Asn Thr Pro
        435                 440                 445
Ala Lys Pro Pro Pro Thr Ile Glu His Gly His Gln Asn Gln Thr Leu
    450                 455                 460
Met Val Gly Ser Ser Ala Ile Leu Pro Cys Gln Ala Ser Gly Lys Pro
465                 470                 475                 480
Thr Pro Gly Ile Ser Trp Leu Arg Asp Gly Leu Pro Ile Asp Ile Thr
                485                 490                 495
Asp Ser Arg Ile Ser Gln His Ser Thr Gly Ser Leu His Ile Ala Asp
            500                 505                 510
Leu Lys Lys Pro Asp Thr Gly Val Tyr Thr Cys Ile Ala Lys Asn Glu
        515                 520                 525
Asp Gly Glu Ser Thr Trp Ser Ala Ser Leu Thr Val Glu Asp His Thr
    530                 535                 540
Ser Asn Ala Gln Phe Val Arg Met Pro Asp Pro Ser Asn Phe Pro Ser
545                 550                 555                 560
Ser Pro Thr Gln Pro Ile Ile Val Asn Val Thr Asp Thr Glu Val Glu
                565                 570                 575
Leu His Trp Asn Ala Pro Ser Thr Ser Gly Ala Gly Pro Ile Thr Gly
            580                 585                 590
Tyr Ile Ile Gln Tyr Tyr Ser Pro Asp Leu Gly Gln Thr Trp Phe Asn
        595                 600                 605
Ile Pro Asp Tyr Val Ala Ser Thr Glu Tyr Arg Ile Lys Gly Leu Lys
    610                 615                 620
Pro Ser His Ser Tyr Met Phe Val Ile Arg Ala Glu Asn Glu Lys Gly
625                 630                 635                 640
Ile Gly Thr Pro Ser Val Ser Ser Ala Leu Val Thr Thr Ser Lys Pro
                645                 650                 655
Ala Ala Gln Val Ala Leu Ser Asp Lys Asn Lys Met Asp Met Ala Ile
            660                 665                 670
Ala Glu Lys Arg Leu Thr Ser Glu Gln Leu Ile Lys Leu Glu Glu Val
        675                 680                 685
Lys Thr Ile Asn Ser Thr Ala Val Arg Leu Phe Trp Lys Lys Arg Lys
    690                 695                 700
Leu Glu Glu Leu Ile Asp Gly Tyr Tyr Ile Lys Trp Arg Gly Pro Pro
705                 710                 715                 720
Arg Thr Asn Asp Asn Gln Tyr Val Asn Val Thr Ser Pro Ser Thr Glu
                725                 730                 735
Asn Tyr Val Val Ser Asn Leu Met Pro Phe Thr Asn Tyr Glu Phe Phe
            740                 745                 750
Val Ile Pro Tyr His Ser Gly Val His Ser Ile His Gly Ala Pro Ser
        755                 760                 765
Asn Ser Met Asp Val Leu Thr Ala Glu Ala Pro Pro Ser Leu Pro Pro
    770                 775                 780
Glu Asp Val Arg Ile Arg Met Leu Asn Leu Thr Thr Leu Arg Ile Ser
785                 790                 795                 800
Trp Lys Ala Pro Lys Ala Asp Gly Ile Asn Gly Ile Leu Lys Gly Phe
                805                 810                 815
Gln Ile Val Ile Val Gly Gln Ala Pro Asn Asn Asn Arg Asn Ile Thr
            820                 825                 830
Thr Asn Glu Arg Ala Ala Ser Val Thr Leu Phe His Leu Val Thr Gly
        835                 840                 845
Met Thr Tyr Lys Ile Arg Val Ala Ala Arg Ser Asn Gly Gly Val Gly
    850                 855                 860
Val Ser His Gly Thr Ser Glu Val Ile Met Asn Gln Asp Thr Leu Glu
865                 870                 875                 880
Lys His Leu Ala Ala Gln Gln Glu Asn Glu Ser Phe Leu Tyr Gly Leu
                885                 890                 895
Ile Asn Lys Ser His Val Pro Val Ile Val Ile Val Ala Ile Leu Ile
            900                 905                 910
Ile Phe Val Val Ile Ile Ile Ala Tyr Cys Tyr Trp Arg Asn Ser Arg
        915                 920                 925
Asn Ser Asp Gly Lys Asp Arg Ser Phe Ile Lys Ile Asn Asp Gly Ser
    930                 935                 940
Val His Met Ala Ser Asn Asn Leu Trp Asp Val Ala Gln Asn Pro Asn
945                 950                 955                 960
Gln Asn Pro Met Tyr Asn Thr Ala Gly Arg Met Thr Met Asn Asn Arg
                965                 970                 975
Asn Gly Gln Ala Leu Tyr Ser Leu Thr Pro Asn Ala Gln Asp Phe Phe
            980                 985                 990
Asn Asn Cys Asp Asp Tyr Ser Gly Thr Met His Arg Pro Gly Ser Glu
        995                 1000                1005
His His Tyr His Tyr Ala Gln Leu Thr Gly Gly Pro Gly Asn Ala Met
    1010                1015                1020
Ser Thr Phe Tyr Gly Asn Gln Tyr His Asp Asp Pro Ser Pro Tyr Ala
1025                1030                1035                1040
Thr Thr Thr Leu Val Leu Ser Asn Gln Gln Pro Ala Trp Leu Asn Asp
                1045                1050                1055
Lys Met Leu Arg Ala Pro Ala Met Pro Thr Asn Pro Val Pro Pro Glu
            1060                1065                1070
Pro Pro Ala Arg Tyr Ala Asp His Thr Ala Gly Arg Arg Ser Arg Ser
        1075                1080                1085
Ser Arg Ala Ser Asp Gly Arg Gly Thr Leu Asn Gly Gly Leu His His
    1090                1095                1100
Arg Thr Ser Gly Ser Gln Arg Ser Asp Ser Pro Pro His Thr Asp Val
1105                1110                1115                1120
Ser Tyr Val Gln Leu His Ser Ser Asp Gly Thr Gly Ser Ser Lys Glu
                1125                1130                1135
Arg Thr Gly Glu Arg Arg Thr Pro Pro Asn Lys Thr Leu Met Asp Phe
            1140                1145                1150
Ile Pro Pro Pro Pro Ser Asn Pro Pro Pro Pro Gly Gly His Val Tyr
        1155                1160                1165
Asp Thr Ala Thr Arg Arg Gln Leu Asn Arg Gly Ser Thr Pro Arg Glu
    1170                1175                1180
Asp Thr Tyr Asp Ser Val Ser Asp Gly Ala Phe Ala Arg Val Asp Val
1185                1190                1195                1200
Asn Ala Arg Pro Thr Ser Arg Asn Arg Asn Leu Gly Gly Arg Pro Leu
                1205                1210                1215
Lys Gly Lys Arg Asp Asp Asp Ser Gln Arg Ser Ser Leu Met Met Asp
            1220                1225                1230
Asp Asp Gly Gly Ser Ser Glu Ala Asp Gly Glu Asn Ser Glu Gly Asp
        1235                1240                1245
Val Pro Arg Gly Gly Val Arg Lys Ala Val Pro Arg Met Gly Ile Ser
    1250                1255                1260
Ala Ser Thr Leu Ala His Ser Cys Tyr Gly Thr Asn Gly Thr Ala Gln
1265                1270                1275                1280
Arg Phe Arg Ser Ile Pro Arg Asn Asn Gly Ile Val Thr Gln Glu Gln
                1285                1290                1295
Thr 
           
             18 
             1651 
             PRT 
             human 
           
            18
Met Lys Trp Lys His Val Pro Phe Leu Val Met Ile Ser Leu Leu Ser
1               5                   10                  15
Leu Ser Pro Asn His Leu Phe Leu Ala Gln Leu Ile Pro Asp Pro Glu
            20                  25                  30
Asp Val Glu Arg Gly Asn Asp His Gly Thr Pro Ile Pro Thr Ser Asp
        35                  40                  45
Asn Asp Asp Asn Ser Leu Gly Tyr Thr Gly Ser Arg Leu Arg Gln Glu
    50                  55                  60
Asp Phe Pro Pro Arg Ile Val Glu His Pro Ser Asp Leu Ile Val Ser
65                  70                  75                  80
Lys Gly Glu Pro Ala Thr Leu Asn Cys Lys Ala Glu Gly Arg Pro Thr
                85                  90                  95
Pro Thr Ile Glu Trp Tyr Lys Gly Gly Glu Arg Val Glu Thr Asp Lys
            100                 105                 110
Asp Asp Pro Arg Ser His Arg Met Leu Leu Pro Ser Gly Ser Leu Phe
        115                 120                 125
Phe Leu Arg Ile Val His Gly Arg Lys Ser Arg Pro Asp Glu Gly Val
    130                 135                 140
Tyr Val Cys Val Ala Arg Asn Tyr Leu Gly Glu Ala Val Ser His Asn
145                 150                 155                 160
Ala Ser Leu Glu Val Ala Ile Leu Arg Asp Asp Phe Arg Gln Asn Pro
                165                 170                 175
Ser Asp Val Met Val Ala Val Gly Glu Pro Ala Val Met Glu Cys Gln
            180                 185                 190
Pro Pro Arg Gly His Pro Glu Pro Thr Ile Ser Trp Lys Lys Asp Gly
        195                 200                 205
Ser Pro Leu Asp Asp Lys Asp Glu Arg Ile Thr Ile Arg Gly Gly Lys
    210                 215                 220
Leu Met Ile Thr Tyr Thr Arg Lys Ser Asp Ala Gly Lys Tyr Val Cys
225                 230                 235                 240
Val Gly Thr Asn Met Val Gly Glu Arg Glu Ser Glu Val Ala Glu Leu
                245                 250                 255
Thr Val Leu Glu Arg Pro Ser Phe Val Lys Arg Pro Ser Asn Leu Ala
            260                 265                 270
Val Thr Val Asp Asp Ser Ala Glu Phe Lys Cys Glu Ala Arg Gly Asp
        275                 280                 285
Pro Val Pro Thr Val Arg Trp Arg Lys Asp Asp Gly Glu Leu Pro Lys
    290                 295                 300
Ser Arg Tyr Glu Ile Arg Asp Asp His Thr Leu Lys Ile Arg Lys Val
305                 310                 315                 320
Thr Ala Gly Asp Met Gly Ser Tyr Thr Cys Val Ala Glu Asn Met Val
                325                 330                 335
Gly Lys Ala Glu Ala Ser Ala Thr Leu Thr Val Gln Glu Pro Pro His
            340                 345                 350
Phe Val Val Lys Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val
        355                 360                 365
Thr Phe Gln Cys Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp
    370                 375                 380
Arg Arg Glu Gly Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln
385                 390                 395                 400
Ser Ser Ser Arg Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr
                405                 410                 415
Asn Val Gln Arg Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn
            420                 425                 430
Val Ala Gly Ser Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val
        435                 440                 445
Ile Ala Asp Arg Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln
    450                 455                 460
Thr Val Ala Val Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly
465                 470                 475                 480
Ser Pro Val Pro Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser
                485                 490                 495
Thr Gln Asp Ser Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile
            500                 505                 510
Arg Tyr Ala Lys Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser
        515                 520                 525
Thr Pro Ser Gly Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu
    530                 535                 540
Phe Gly Val Pro Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile
545                 550                 555                 560
Pro Ser Ala Pro Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr
                565                 570                 575
Val Thr Leu Ser Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr
            580                 585                 590
Ser Tyr Ile Ile Glu Ala Phe Ser His Ala Ser Gly Ser Ser Trp Gln
        595                 600                 605
Thr Val Ala Glu Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu
    610                 615                 620
Lys Pro Asn Ala Ile Tyr Leu Phe Leu Val Arg Ala Ala Asn Ala Tyr
625                 630                 635                 640
Gly Ile Ser Asp Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp
                645                 650                 655
Val Leu Pro Thr Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu
            660                 665                 670
Leu Gly Asn Ala Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser
        675                 680                 685
Ser Ser Ile Glu Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile
    690                 695                 700
Gln Gly Tyr Lys Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu
705                 710                 715                 720
Ser Asp Trp Leu Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val
                725                 730                 735
Val Ile Pro Asp Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg
            740                 745                 750
Pro Phe Phe Asn Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala
        755                 760                 765
Lys Thr Leu Glu Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val
    770                 775                 780
Ser Lys Asn Asp Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro
785                 790                 795                 800
Pro Pro Glu Asp Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp
                805                 810                 815
Cys Leu Gly Asn Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly
            820                 825                 830
Ser Thr Phe Ser Val Val Ile Pro Phe Leu Val Pro Gly Ile Arg Tyr
        835                 840                 845
Ser Val Glu Val Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser
    850                 855                 860
Glu Pro Gln Phe Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro
865                 870                 875                 880
Glu Asp Gln Val Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln
                885                 890                 895
Pro Ala Phe Ile Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met
            900                 905                 910
Val Phe Ser Ile Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu
        915                 920                 925
Thr Ser Thr Tyr Ala Gly Ile Arg Lys Val Pro Ser Phe Thr Phe Thr
    930                 935                 940
Pro Thr Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser Ser Gly Gly
945                 950                 955                 960
Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln Pro Trp Leu
                965                 970                 975
Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp Cys Ser Ile
            980                 985                 990
Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn Leu Thr Thr
        995                 1000                1005
Tyr Ser Arg Pro Ala Asp Cys Ile Ala Asn Tyr Asn Asn Gln Leu Asp
    1010                1015                1020
Asn Lys Gln Thr Asn Leu Met Leu Pro Glu Ser Thr Val Tyr Gly Asp
1025                1030                1035                1040
Val Asp Leu Ser Asn Lys Ile Asn Glu Met Lys Thr Phe Asn Ser Pro
                1045                1050                1055
Asn Leu Lys Asp Gly Arg Phe Val Asn Pro Ser Gly Gln Pro Thr Pro
            1060                1065                1070
Tyr Ala Thr Thr Gln Leu Ile Gln Ser Asn Leu Ser Asn Asn Met Asn
        1075                1080                1085
Asn Gly Ser Gly Asp Ser Gly Glu Lys His Trp Lys Pro Leu Gly Gln
    1090                1095                1100
Gln Lys Gln Glu Val Ala Pro Val Gln Tyr Asn Ile Val Glu Gln Asn
1105                1110                1115                1120
Lys Leu Asn Lys Asp Tyr Arg Ala Asn Asp Thr Val Pro Pro Thr Ile
                1125                1130                1135
Pro Tyr Asn Gln Ser Tyr Asp Gln Asn Thr Gly Gly Ser Tyr Asn Ser
            1140                1145                1150
Ser Asp Arg Gly Ser Ser Thr Ser Gly Ser Gln Gly His Lys Lys Gly
        1155                1160                1165
Ala Arg Thr Pro Lys Val Pro Lys Gln Gly Gly Met Asn Trp Ala Asp
    1170                1175                1180
Leu Leu Pro Pro Pro Pro Ala His Pro Pro Pro His Ser Asn Ser Glu
1185                1190                1195                1200
Glu Tyr Asn Ile Ser Val Asp Glu Ser Tyr Asp Gln Glu Met Pro Cys
                1205                1210                1215
Pro Val Pro Pro Ala Arg Met Tyr Leu Gln Gln Asp Glu Leu Glu Glu
            1220                1225                1230
Glu Glu Asp Glu Arg Gly Pro Thr Pro Pro Val Arg Gly Ala Ala Ser
        1235                1240                1245
Ser Pro Ala Ala Val Ser Tyr Ser His Gln Ser Thr Ala Thr Leu Thr
    1250                1255                1260
Pro Ser Pro Gln Glu Glu Leu Gln Pro Met Leu Gln Asp Cys Pro Glu
1265                1270                1275                1280
Glu Thr Gly His Met Gln His Gln Pro Asp Arg Arg Arg Gln Pro Val
                1285                1290                1295
Ser Pro Pro Pro Pro Pro Arg Pro Ile Ser Pro Pro His Thr Tyr Gly
            1300                1305                1310
Tyr Ile Ser Gly Pro Leu Val Ser Asp Met Asp Thr Asp Ala Pro Glu
        1315                1320                1325
Glu Glu Glu Asp Glu Ala Asp Met Glu Val Ala Lys Met Gln Thr Arg
    1330                1335                1340
Arg Leu Leu Leu Arg Gly Leu Glu Gln Thr Pro Ala Ser Ser Val Gly
1345                1350                1355                1360
Asp Leu Glu Ser Ser Val Thr Gly Ser Met Ile Asn Gly Trp Gly Ser
                1365                1370                1375
Ala Ser Glu Glu Asp Asn Ile Ser Ser Gly Arg Ser Ser Val Ser Ser
            1380                1385                1390
Ser Asp Gly Ser Phe Phe Thr Asp Ala Asp Phe Ala Gln Ala Val Ala
        1395                1400                1405
Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala Arg Arg Gln Met Gln
    1410                1415                1420
Asp Ala Ala Gly Arg Arg His Phe His Ala Ser Gln Cys Pro Arg Pro
1425                1430                1435                1440
Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser Ala Ala Val Met Gln
                1445                1450                1455
Lys Thr Arg Pro Ala Lys Lys Leu Lys His Gln Pro Gly His Leu Arg
            1460                1465                1470
Arg Glu Thr Tyr Thr Asp Asp Leu Pro Pro Pro Pro Val Pro Pro Pro
        1475                1480                1485
Ala Ile Lys Ser Pro Thr Ala Gln Ser Lys Thr Gln Leu Glu Val Arg
    1490                1495                1500
Pro Val Val Val Pro Lys Leu Pro Ser Met Asp Ala Arg Thr Asp Arg
1505                1510                1515                1520
Ser Ser Asp Arg Lys Gly Ser Ser Tyr Lys Gly Arg Glu Val Leu Asp
                1525                1530                1535
Gly Arg Gln Val Val Asp Met Arg Thr Asn Pro Gly Asp Pro Arg Glu
            1540                1545                1550
Ala Gln Glu Gln Gln Asn Asp Gly Lys Gly Arg Gly Asn Lys Ala Ala
        1555                1560                1565
Lys Arg Asp Leu Pro Pro Ala Lys Thr His Leu Ile Gln Glu Asp Ile
    1570                1575                1580
Leu Pro Tyr Cys Arg Pro Thr Phe Pro Thr Ser Asn Asn Pro Arg Asp
1585                1590                1595                1600
Pro Ser Ser Ser Ser Ser Met Ser Ser Arg Gly Ser Gly Ser Arg Gln
                1605                1610                1615
Arg Glu Gln Ala Asn Val Gly Arg Arg Asn Ile Ala Glu Met Gln Val
            1620                1625                1630
Leu Gly Gly Tyr Glu Arg Gly Glu Asp Asn Asn Glu Glu Leu Glu Glu
        1635                1640                1645
Thr Glu Ser
    1650 
           
             19 
             434 
             PRT 
             human 
             
               misc_feature 
               (285)..(396) 
               note=“Xaa signifies gap in sequence” 
             
           
            19
Gln Ile Val Ala Gln Gly Arg Thr Val Thr Phe Pro Cys Glu Thr Lys
1               5                   10                  15
Gly Asn Pro Gln Pro Ala Val Phe Trp Gln Lys Glu Gly Ser Gln Asn
            20                  25                  30
Leu Leu Phe Pro Asn Gln Pro Gln Gln Pro Asn Ser Arg Cys Ser Val
        35                  40                  45
Ser Pro Thr Gly Asp Leu Thr Ile Thr Asn Ile Gln Arg Ser Asp Ala
    50                  55                  60
Gly Tyr Tyr Ile Cys Gln Ala Leu Thr Val Ala Gly Ser Ile Leu Ala
65                  70                  75                  80
Lys Ala Gln Leu Glu Val Thr Asp Val Leu Thr Asp Arg Pro Pro Pro
                85                  90                  95
Ile Ile Leu Gln Gly Pro Ala Asn Gln Thr Leu Ala Val Asp Gly Thr
            100                 105                 110
Ala Leu Leu Lys Cys Lys Ala Thr Gly Asp Pro Leu Pro Val Ile Ser
        115                 120                 125
Trp Leu Lys Glu Gly Phe Thr Phe Pro Gly Arg Asp Pro Arg Ala Thr
    130                 135                 140
Ile Gln Glu Gln Gly Thr Leu Gln Ile Lys Asn Leu Arg Ile Ser Asp
145                 150                 155                 160
Thr Gly Thr Tyr Thr Cys Val Ala Thr Ser Ser Ser Gly Glu Ala Ser
                165                 170                 175
Trp Ser Ala Val Leu Asp Val Thr Glu Ser Gly Ala Thr Ile Ser Lys
            180                 185                 190
Asn Tyr Asp Leu Ser Asp Leu Pro Gly Pro Pro Ser Lys Pro Gln Val
        195                 200                 205
Thr Asp Val Thr Lys Asn Ser Val Thr Leu Ser Trp Gln Pro Gly Thr
    210                 215                 220
Pro Gly Thr Leu Pro Ala Ser Ala Tyr Ile Ile Glu Ala Phe Ser Gln
225                 230                 235                 240
Ser Val Ser Asn Ser Trp Gln Thr Val Ala Asn His Val Lys Thr Thr
                245                 250                 255
Leu Tyr Thr Val Arg Gly Leu Arg Pro Asn Thr Ile Tyr Leu Phe Met
            260                 265                 270
Val Arg Ala Ile Asn Pro Lys Val Ser Val Thr Gln Xaa Lys Pro Gln
        275                 280                 285
Lys Asn Asn Gly Ser Thr Trp Ala Asn Val Pro Leu Pro Pro Pro Pro
    290                 295                 300
Val Gln Pro Leu Pro Gly Thr Glu Leu Glu His Tyr Ala Val Glu Gln
305                 310                 315                 320
Gln Glu Asn Gly Tyr Asp Ser Asp Ser Trp Cys Pro Pro Leu Pro Val
                325                 330                 335
Gln Thr Tyr Leu His Gln Gly Leu Glu Asp Glu Leu Glu Glu Asp Asp
            340                 345                 350
Asp Arg Val Pro Thr Pro Pro Val Arg Gly Val Ala Ser Ser Pro Ala
        355                 360                 365
Ile Ser Phe Gly Gln Gln Ser Thr Ala Thr Leu Thr Pro Ser Pro Arg
    370                 375                 380
Glu Glu Met Gln Pro Met Leu Gln Ala Ser Pro Xaa Phe Thr Ser Ser
385                 390                 395                 400
Gln Arg Pro Arg Pro Thr Ser Pro Phe Ser Thr Asp Ser Asn Thr Ser
                405                 410                 415
Ala Ala Leu Ser Gln Ser Gln Arg Pro Arg Pro Thr Lys Lys His Lys
            420                 425                 430
Gly Gly 
           
             20 
             148 
             PRT 
             mouse 
           
            20
Ala Gln Ala Val Ala Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala
1               5                   10                  15
Arg Arg Gln Met Gln Asp Ala Ala Gly Arg Arg His Phe His Ala Ser
            20                  25                  30
Gln Cys Pro Arg Pro Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser
        35                  40                  45
Ala Val Val Ile Gln Lys Ala Arg Pro Ala Lys Lys Gln Lys His Gln
    50                  55                  60
Pro Gly His Leu Arg Arg Glu Ala Tyr Ala Asp Asp Leu Pro Pro Pro
65                  70                  75                  80
Pro Val Pro Pro Pro Ala Ile Lys Ser Pro Thr Val Gln Ser Lys Ala
                85                  90                  95
Gln Leu Glu Val Arg Pro Val Met Val Pro Lys Leu Ala Ser Ile Glu
            100                 105                 110
Ala Arg Thr Asp Arg Ser Ser Asp Arg Lys Gly Gly Ser Tyr Lys Gly
        115                 120                 125
Arg Glu Ala Leu Asp Gly Arg Gln Val Thr Asp Leu Arg Thr Asn Pro
    130                 135                 140
Ser Asp Pro Arg
145