Novel Metalloproteinases

Abstract

The present invention relates to novel metalloproteinase- like proteins. In particular, isolated nucleic acid molecules are provided encoding the human TACE-like and matrilysin-like proteins. TACE-like and matrilysin-like polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of TACE-like and matrilysin-like activity. Also provided are diagnostic methods for detecting cancer and therapeutic methods for cancer and other disorders characterized by an over or under production of these metalloproteinases.

1 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 261 in SEQ ID NO:5;    (b) a nucleotide sequence encoding a polypeptide comprising amino acids from about 2 to about 261 in SEQ ID NO:5;    (c) a nucleotide sequence encoding a polypeptide comprising amino acids from about 23 to about 261 in SEQ ID NO:5;    (d) a nucleotide sequence encoding a polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209055;    (e) a nucleotide sequence encoding the mature matrilysin-like polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209055; and    (f) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), or (e).    
     
     
         2 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in SEQ ID NO:4.  
     
     
         3 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in SEQ ID NO:4 encoding the matrilysin-like polypeptide having the amino acid sequence in SEQ ID NO:5.  
     
     
         4 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in SEQ ID NO:4 encoding the mature matrilysin-like polypeptide having the amino acid sequence in SEQ ID NO:5.  
     
     
         5 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence of the cDNA clone contained in ATCC™ Deposit No. 209055.  
     
     
         6 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the matrilysin-like polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209055.  
     
     
         7 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the mature matrilysin-like polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209055.  
     
     
         8 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), (e), or (f) of  claim 1  wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.  
     
     
         9 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a matrilysin-like polypeptide having an amino acid sequence in (a), (b), (c), (d), or (e) of  claim 1 .  
     
     
         10 . The isolated nucleic acid molecule of  claim 9 , which encodes an epitope-bearing portion of a matrilysin-like polypeptide selected from the group consisting of: a polypeptide comprising amino acid residues from about 24 to about 71 in SEQ ID NO:5; a polypeptide comprising amino acid residues from about 81 to about 125 in SEQ ID NO:5; a polypeptide comprising amino acid residues from about 141 to about 167 in SEQ ID NO:5; a polypeptide comprising amino acid residues from about 178 to about 202 in SEQ ID NO:5;and a polypeptide comprising amino acid residues from about 212 to about 260 in SEQ ID NO:5.  
     
     
         11 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
     
     
         12 . A recombinant vector produced by the method of  claim 11 .  
     
     
         13 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 12  into a host cell.  
     
     
         14 . A recombinant host cell produced by the method of  claim 13 .  
     
     
         15 . A recombinant method for producing a matrilysin-like polypeptide, comprising culturing the recombinant host cell of  claim 14  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         16 . An isolated matrilysin-like polypeptide having an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) amino acids from about 1 to about 261 in SEQ ID NO:5;    (b) amino acids from about 2 to about 261 in SEQ ID NO:5;    (c) amino acids from about 23 to about 261 in SEQ ID NO:5;    (d) the amino acid sequence of the matrilysin-like polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209055;    (e) the amino acid sequence of the mature matrilysin-like polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 209055; and    (f) the amino acid sequence of an epitope-bearing portion of any one of the polypeptides of (a), (b), (c), (d), or (e).    
     
     
         17 . The isolated polypeptide of  claim 16 , which is produced or contained in a recombinant host cell.  
     
     
         18 . An isolated antibody or fragment thereof that specifically binds the polypeptide of  claim 16 .  
     
     
         19 . A method of detecting the matrilysin-like polypeptide in a biological sample comprising: 
 (a) contacting the biological sample with an antibody or fragment thereof that specifically binds a protein consisting of amino acid residues 1 to about 261 in SEQ ID NO:5; and    (b) detecting said matrilysin-like polypeptide in the biological sample bound to said antibody or fragment thereof.