Mch2, an apoptotic cysteine protease, and compositions for making and methods of using the same

Abstract

A substantially pure protein that is a member of the apoptotic Ced-3/Ice cysteine protease gene family, Mch2α, and an inactive isoform of it, Mch2β, are disclosed. Isolated nucleic acid molecules that encode Mch2α and Mch2β, respectively, are disclosed. Pharmaceutical compositions comprising a pharmaceutically acceptable carrier in combination with the protein or the nucleic acid molecules are disclosed. Fragments of nucleic acid molecules that encode Mch2α and Mch2β having at least 10 nucleotides and oligonucleotide molecule comprising a nucleotide sequence complimentary to a nucleotide sequence of at least 10 nucleotides are disclosed. Recombinant expression vectors that comprise the nucleic acid molecule that encode Mch2α or Mch2β, and host cells that comprise such recombinant vectors are disclosed. Antibodies that bind to an epitope on Mch2α and/or Mch2β are disclosed. Methods of identifying inhibitors, activators and substrates of Mch2α are disclosed. Antisense compounds and methods of using the same are disclosed.

1 . A substantially pure protein having the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 7.  
     
     
         2 . The protein of  claim 1  wherein said protein has the amino acid sequence of SEQ ID NO: 5.  
     
     
         3 . The protein of  claim 1  wherein said protein has the amino acid sequence of SEQ ID NO: 7.  
     
     
         4 . A pharmaceutical composition comprising the protein of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         5 . An isolated nucleic acid molecule that comprises a nucleic acid sequence that encodes the protein of  claim 1 .  
     
     
         6 . A pharmaceutical composition comprising the nucleic acid molecule of  claim 5  and a pharmaceutically acceptable carrier.  
     
     
         7 . An isolated nucleic acid molecule consisting of SEQ ID NO: 4 or SEQ ID NO: 5 or a fragment thereof having at least 10 nucleotides.  
     
     
         8 . The nucleic acid molecule of  claim 7  consisting of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         9 . A recombinant expression vector comprising the nucleic acid molecule of  claim 8 .  
     
     
         10 . A host cell comprising the recombinant expression vector of  claim 9 .  
     
     
         11 . The nucleic acid molecule of  claim 7  consisting of a fragment of SEQ ID NO: 4 or SEQ ID NO: 6 having at least 10 nucleotides.  
     
     
         12 . The nucleic acid molecule of  claim 7  consisting of a fragment of SEQ ID NO: 4 or SEQ ID NO: 6 having 12-150 nucleotides.  
     
     
         13 . The nucleic acid molecule of  claim 7  consisting of a fragment of SEQ ID NO: 4 or SEQ ID NO: 6 having 15-50 nucleotides.  
     
     
         14 . An oligonucleotide molecule comprising a nucleotide sequence complimentary to a nucleotide sequence of at least 5 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         15 . The oligonucleotide molecule of  claim 14  wherein said oligonucleotide molecule comprises a nucleotide sequence complimentary to a nucleotide sequence of 5-50 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         16 . The oligonucleotide molecule of  claim 14  wherein said oligonucleoride molecule comprises a nuclectide sequence complimentary to a nucleotide sequence of 10-40 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         17 . The oligonucleotide molecule of  claim 14  wherein said oligonucleotide molecule comprises a nucleotide sequence complimentary to a nucleotide sequence of 15-25 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         18 . The oligcnucleotide molecule of  claim 14  consisting of a nucleotide sequence complimentary to a nucleotide sequence of at least 10-150 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         19 . The oligonucleotide molecule of  claim 18  consisting of a nucleotide sequence complimentary to a nucleotide sequence of at least 18-28 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.  
     
     
         20 . An isolated antibody which binds to an epitope on SEQ ID NO: 5 and/or SEQ ID NO: 7.  
     
     
         21 . The antibody of  claim 20  wherein said antibody is a monoclonal antibody.  
     
     
         22 . A method of identifying inhibitors of Mch2α activity comprising the steps of: 
 performing a test assay by contacting an Mch2α with a substrate in the presence of a test compound under conditions in which said Mch2α processes said substrate in the absence of said test compound,  
 determining whether said substrate is processed.  
 
     
     
         23 . A method of identifying activators of Mch2α activity comprising the steps of: 
 performing a test assay by contacting an Mch2α with a substrate in the presence of a test compound under conditions in which said Mch2α processes said substrate in the absence of said test compound,  
 comparing the rate of processing of the substrate in the presence of the Best compound to the rate of processing the substrate in the absence of the compound.  
 
     
     
         24 . A method of inhibiting expression of Mch2 comprising the step of: 
 contacting cells that express Mch2 with a nucleic acid molecule that comprises oligonucleotide molecule that comprises a nucleotide sequence complimentary to a nucleotide sequence of 5-50 nucleotides of SEQ ID NO: 4 or SEQ ID NO: 6.