Human cDNAs encoding polypeptides having kinase functions

Abstract

The invention is directed to purified and isolated human polypeptides having kinase function, the nucleic acids encoding such polypeptides, processes for production of recombinant forms of such polypeptides, antibodies generated against these polypeptides, fragmented peptides derived from these polypeptides, the use of such polypeptides and fragmented peptides in phosphorylation reactions and as molecular weight markers, the use of such polypeptides and fragmented peptides as controls for peptide fragmentation, the use of such polypeptides in screening assays, and kits comprising these reagents.

1 . An isolated polynucleotide molecule selected from: 
 a) a nucleotide sequence encoding a polypeptide having kinase activity comprising a sequence having at least 80% identity to the polypeptide sequence of SEQ ID NO:9;    b) a nucleotide sequence encoding a polypeptide having kinase activity comprising a sequence having at least 90% identity to the polypeptide of SEQ ID NO:9;    c) degenerate nucleotide sequences encoding a polypeptide of a) or b);    d) the nucleotide sequence of SEQ ID NO:3;    e) nucleotide sequences capable of hybridization to nucleotide of a) or b) under conditions of moderate stringency;    f) nucleotide sequences complementary to a), b), c), d) or e).    
     
     
         2 . An isolated polynucleotide molecule encoding a polypeptide comprising the sequence of SEQ ID NO:9.  
     
     
         3 . A recombinant vector that directs the expression of the polynucleotide molecule of  claim 2 .  
     
     
         4 . An isolated polypeptide having kinase activity encoded by the polynucleotide molecule of  claim 1 .  
     
     
         5 . An isolated polypeptide according to  claim 4  having a molecular weight of approximately 39,284 Daltons as determined by SDS-PAGE.  
     
     
         6 . An isolated polypeptide according to  claim 4  in non-glycosylated form.  
     
     
         7 . A recombinant host cell comprising the polynucleotide of  claim 2 .  
     
     
         8 . A method for the production of a polypeptide encoded by the polynucleotide of  claim 2  comprising culturing a recombinant host cell comprising the polynucleotide of  claim 2  under conditions promoting expression of said polypeptide.  
     
     
         9 . The method of  claim 8 , wherein the host cell is selected from the group consisting of bacterial cells, yeast cells, plant cells, insect cells, and animal cells.  
     
     
         10 . An isolated polypeptide having kinase activity produced by the method of  claim 8 .  
     
     
         11 . An isolated polypeptide having kinase activity selected from: 
 a) a polypeptide comprising a sequence having at least 80% identity to the polypeptide sequence of SEQ ID NO:9;    b) a polypeptide comprising a sequence having at least 80% identity to the polypeptide sequence of SEQ ID NO:9;    c) a polypeptide sequence comprising SEQ ID NO:9    
     
     
         12 . Isolated antibodies that bind to a polypeptide of  claim 11 .  
     
     
         13 . A method of screening a candidate molecule to identify its ability to inhibit (antagonize) or agonize a polypeptide encoded by a polynucleotide of  claim 2 , said method comprising the steps of: 
 a) adding said candidate molecule to a medium which contains cells expression said polypeptide;    b) determining the level of biological activity of said polypeptide in said medium; and    c) comparing the level of biological activity of step (b) with the level of biological activity that occurs in said medium in the absence of said candidate molecule;    wherein a decreased level of biological activity of step (b) as compared to the level of biological activity that occurs in said medium in the absence of said candidate molecule indicates an antagonist and an increased level of biological activity indicates an agonist.    
     
     
         14 . A method of screening a candidate molecule to identify its ability to mimic the biological activity of a polypeptide encoded by a polynucleotide of  claim 2 , said method comprising the steps of: 
 a) adding said candidate molecule to a biological assay to determine its biological effects; and    b) comparing said biological effects to said candidate molecule with the biological effects of said polypeptide.    
     
     
         15 . A method for screening a candidate molecule for its ability to bind to a polypeptide, the method comprising determining if said candidate molecule binds to a polypeptide of  claim 11.