Polynucleotide encoding a novel cysteine protease of the calpain superfamily, CAN-12, and variants thereof

Abstract

The present invention provides novel polynucleotides encoding CAN-12 polypeptides, fragments and homologues thereof. The present invention also provides polynucleotides encoding variants of CAN-12 polypeptides, CAN-12v1 and CAN-12v2. Also provided are vectors, host cells, antibodies, and recombinant and synthetic methods for producing said polypeptides. The invention further relates to diagnostic and therapeutic methods for applying these novel CAN-12, CAN-12v1, and CAN-12v2 polypeptides to the diagnosis, treatment, and/or prevention of various diseases and/or disorders related to these polypeptides, particularly neuro- and musculo-degenerative conditions. The invention further relates to screening methods for identifying agonists and antagonists of the polynucleotides and polypeptides of the present invention.

1 - 23 . (canceled)  
     
     
         24 . An isolated polypeptide comprising a polypeptide sequence selected from the group consisting of: 
 (a) an isolated polypeptide comprising amino acids 1 to 697 of SEQ ID NO:56;    (b) an isolated polypeptide comprising amino acids 2 to 697 of SEQ ID NO:56;    (c) an isolated polypeptide comprising a polypeptide sequence encoded by nucleotides 9 to 2099 of SEQ ID NO:55; and    (d) an isolated polypeptide comprising a polypeptide sequence encoded by nucleotides 12 to 2099 of SEQ ID NO:55.    
     
     
         25 . The isolated polypeptide of  claim 24 , wherein said polypeptide is (a).  
     
     
         26 . The isolated polypeptide of  claim 24 , wherein said polypeptide is (b).  
     
     
         27 . The isolated polypeptide of  claim 24 , wherein said polypeptide is (c).  
     
     
         28 . The isolated polypeptide of  claim 24 , wherein said polypeptide is (d).  
     
     
         29 . An isolated polypeptide produced by a method comprising: 
 (a) culturing an isolated recombinant host cell comprising a vector that comprises the coding region encoding the polypeptide of  claim 24  under conditions such that the polypeptide of  claim 24  is expressed; and    (b) recovering said polypeptide.    
     
     
         30 . The isolated polypeptide of  claim 24  further comprising a heterologous polypeptide sequence.  
     
     
         31 . The isolated polypeptide of  claim 28  wherein said heterologous polypeptide is the Fc domain of immunoglobulin.  
     
     
         32 . An isolated polypeptide comprising a polypeptide sequence selected from the group consisting of: 
 (a) an isolated polypeptide comprising amino acids 1 to 694 of SEQ ID NO:54;    (b) an isolated polypeptide comprising amino acids 2 to 694 of SEQ ID NO:54;    (c) an isolated polypeptide comprising a polypeptide sequence encoded by nucleotides 9 to 2090 of SEQ ID NO:53; and    (d) an isolated polypeptide comprising a polypeptide sequence encoded by nucleotides 12 to 2090 of SEQ ID NO:53.    
     
     
         33 . The isolated polypeptide of  claim 32 , wherein said polypeptide is (a).  
     
     
         34 . The isolated polypeptide of  claim 32 , wherein said polypeptide is (b).  
     
     
         35 . The isolated polypeptide of  claim 32 , wherein said polypeptide is (c).  
     
     
         36 . The isolated polypeptide of  claim 32 ,.wherein said polypeptide is (d).  
     
     
         37 . An isolated polypeptide produced by a method comprising: 
 (a) culturing an isolated recombinant host cell comprising a vector that comprises the coding region encoding the polypeptide of  claim 32  under conditions such that the polypeptide of  claim 32  is expressed; and    (b) recovering said polypeptide..    
     
     
         38 . The isolated polypeptide of  claim 32  further comprising a heterologous polypeptide sequence.  
     
     
         39 . The isolated polypeptide of  claim 38  wherein said heterologous polypeptide is the Fc domain of immunoglobulin.  
     
     
         40 . An isolated polypeptide comprising the polypeptide encoded by the CAN-12v1 or the CAN-12v2 cDNA clone contained in ATCC Deposit No: PTA-3434.  
     
     
         41 . An isolated polypeptide comprising amino acids from position 2 through position 587 of SEQ ID NO:56, wherein said polypeptide has calcium-dependent cysteine protease activity.  
     
     
         42 . An isolated nucleic acid comprising a polynucleotide encoding a polypeptide comprising the sequence of amino acids from position 90 through position 697 of SEQ ID NO:56, wherein said polypeptide has calcium-dependent cysteine protease activity.  
     
     
         43 . An isolated polypeptide comprising amino acids from position 2 through position 584 of SEQ ID NO:54, wherein said polypeptide has calcium-dependent cysteine protease activity.  
     
     
         44 . An isolated nucleic acid comprising a polynucleotide encoding a polypeptide comprising the sequence of amino acids from position 90 through position 694 of SEQ ID NO:54, wherein said polypeptide has calcium-dependent cysteine protease activity.  
     
     
         45 . An isolated polypeptide having as least 95.0% identity to amino acids 2 to 697 of SEQ ID NO:56, wherein percent identity is calculated using a CLUSTALW global sequence alignment using default parameters, wherein said polypeptide has calcium-dependent cysteine protease activity.  
     
     
         46 . An isolated polypeptide having as least 95.0% identity to amino acids 2 to 694 of SEQ ID NO:54, wherein percent identity is calculated using a CLUSTALW global sequence alignment using default parameters, wherein said polypeptide has calcium-dependent cysteine protease activity.