Pancreas-derived plasminogen activator inhibitor

Abstract

The present invention relates to a novel member of the plasminogen activator inhibitor protein family. In particular, isolated nucleic acid molecules are provided encoding the pancreas-derived plasminogen activator inhibitor protein. Pancreas-derived plasminogen activator inhibitor polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to methods for treating physiologic and pathologic disease conditions and diagnostic methods for detecting pathologic disorders.

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 −14 to about 378 in SEQ ID NO:2;    (b) a nucleotide sequence encoding a polypeptide comprising amino acids from about −13 to about 378 in SEQ ID NO:2;    (c) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 378 in SEQ ID NO:2;    (d) a nucleotide sequence encoding a polypeptide comprising amino acids from about −18 to about 387 in SEQ ID NO:13;    (e) a nucleotide sequence encoding a polypeptide comprising amino acids from about −17 to about 387 in SEQ ID NO:13;    (f) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 387 in SEQ ID NO:13;    (g) a nucleotide sequence encoding a polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATTC™ Deposit No. 97657;    (h) a nucleotide sequence encoding the mature PAPAI polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATTC™ Deposit No. 97657; and    (i) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), or (h).    
     
     
         2 . 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), (f), (g), (h), or (i) 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.  
     
     
         3 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a PAPAI polypeptide having an amino acid sequence in (a), (b), (c), (d), (e), (f), or (g) of  claim 1 .  
     
     
         4 . The isolated nucleic acid molecule of  claim 3 , which encodes an epitope-bearing portion of a PAPAI polypeptide selected from the group consisting of: a polypeptide comprising amino acid residues from about 6 to about 16 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 31 to about 36 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 46 to about 76 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 111 to about 121 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 146 to about 161 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 206 to about 211 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 236 to about 246 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 306 to about 316 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 361 to about 366 in SEQ ID NO:2; a polypeptide comprising amino acid residues from about 2 to about 12 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 27 to about 32 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 42 to about 72 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 107 to about 117 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 142 to about 157 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 202 to about 207 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 232 to about 242 in SEQ ID NO:13; a polypeptide comprising amino acid residues from about 302 to about 312 in SEQ ID NO:13; and a polypeptide comprising amino acid residues from about 357 to about 362 in SEQ ID NO:13.  
     
     
         5 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
     
     
         6 . A recombinant vector produced by the method of  claim 5 .  
     
     
         7 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 6  into a host cell.  
     
     
         8 . A recombinant host cell produced by the method of  claim 7 .  
     
     
         9 . A recombinant method for producing a PAPAI polypeptide, comprising culturing the recombinant host cell of  claim 8  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         10 . An isolated PAPAI polypeptide having an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) amino acids from about −14 to about 378 in SEQ ID NO:2;    (b) amino acids from about −13 to about 378 in SEQ ID NO:2;    (c) amino acids from about 1 to about 378 in SEQ ID NO:2;    (d) amino acids from about −18 to about 387 in SEQ ID NO:13;    (e) amino acids from about −17 to about 387 in SEQ ID NO:13;    (f) amino acids from about 1 to about 387 in SEQ ID NO:13;    (g) the amino acid sequence of the PAPAI polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATTC™ Deposit No. 97657;    (h) the amino acid sequence of the mature PAPAI polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATTC™ Deposit No. 97657; and    (i) the amino acid sequence of an epitope-bearing portion of any one of the polypeptides of (a), (b), (c), (d), (e), (f), (g), or (h).    
     
     
         11 . The isolated polypeptide of  claim 10 , which is produced or contained in a recombinant host cell.  
     
     
         12 . The isolated polypeptide of  claim 10 , wherein said recombinant host cell is mammalian.  
     
     
         13 . An isolated antibody that binds specifically to a plasminogen activator inhibitor polypeptide of  claim 10 .  
     
     
         14 . A method for treatment of an individual in need of an increased level of pancreas-derived plasminogen activator inhibitor activity comprising administering to said individual a composition comprising an isolated polypeptide of  claim 10 .  
     
     
         15 . A method for treatment of an individual in need of an increased level of pancreas-derived plasminogen activator inhibitor activity comprising administering to said individual a composition comprising a recombinant vector of  claim 6.