DNA encoding the human serine protease C-E

Abstract

Here we describe the molecular identification of a cDNA encoding a novel serine protease we have termed protease C-E. The deduced amino acid sequence, and it alignment with other well-characterized serine proteases indicates that it is a member of the S1 serine protease family. We have found that the protease C-E mRNA is expressed in pancreas, placenta, prostate, small intestine, stomach, spleen, fibroblasts and epidermis, as well as in certain regions of the brain i.e., cerebellum, cerebral cortex, pituitary and hippocampus. Enzymatically active protease C-E, as produced using the methodologies described herein, is amenable to further biochemical analyses for the identification of physiological substrates and specific modulators.

1 . An isolated and purified nucleic acid molecule that encodes protease C-E, and functional derivatives thereof.  
     
     
         2 . The isolated and purified nucleic acid molecule of  claim 1 , having a nucleotide sequence selected from a group consisting of: (SEQ.ID.NO.:1), (SEQ.ID.NO.:2) and functional derivatives thereof.  
     
     
         3 . The isolated and purified nucleic acid molecule of  claim 1 , wherein said molecule is selected from a group consisting of DNA, genomic DNA, cDNA, RNA and mRNA.  
     
     
         4 . An expression vector for expression of protease C-E protein in a recombinant host, wherein said vector contains a nucleic acid molecule having a nucleotide sequence encoding protease C-E protein, and functional derivatives thereof.  
     
     
         5 . The expression vector of  claim 4 , wherein the expression vector contains a nucleotide sequence encoding protease C-E protein selected from a group consisting of (SEQ.ID.NO.:1), (SEQ.ID.NO.:2) and functional derivatives thereof.  
     
     
         6 . The expression vector of  claim 4 , wherein the nucleic acid molecule encoding protease C-E protein is genomic DNA.  
     
     
         7 . A recombinant host cell containing the expression vector of  claim 4 .  
     
     
         8 . The recombinant host cell of  claim 7 , wherein said expression vector contains a nucleotide sequence selected from a group consisting of (SEQ.ID.NO.:1), (SEQ.ID.NO.:2) and functional derivatives thereof.  
     
     
         9 . The recombinant host cell of  claim 7 , wherein said nucleotide sequence is genomic DNA.  
     
     
         10 . A protein in substantially pure form that functions as protease C-E protein.  
     
     
         11 . The protein according to  claim 10 , having an amino acid sequence selected from a group consisting of (SEQ.ID.NO.:7), (SEQ.ID.NO.:8) and functional derivatives thereof.  
     
     
         12 . A monospecific antibody immunologically reactive with protease C-E protein.  
     
     
         13 . The antibody of  claim 12 , wherein the antibody blocks protease activity of the protease C-E protein.  
     
     
         14 . A process for expression of protease C-E protein in a recombinant host cell, comprising: 
 (a) transferring the expression vector of  claim 4  into suitable host cells; and    (b) culturing the host cells of step (a) under conditions which allow expression of the protease C-E protein from the expression vector.    
     
     
         15 . A method of identifying compounds that modulate protease C-E protein activity, comprising: 
 (a) combining a modulator of protease C-E protein activity, protease C-E protein, and a labeled substrate; and    (b) measuring a change in the labeled substrate.    
     
     
         16 . The method of  claim 15  wherein the labeled substrate is selected from the group consisting of flourogenic, colormetric, radiometric, and fluorescent resonance energy transfer (FRET).  
     
     
         17 . A compound active in the method of  claim 15 , wherein said compound is a modulator of protease C-E serine protease activity.  
     
     
         18 . The compound of  claim 16 , wherein said compound is an agonist or antagonist of protease C-E serine protease activity.  
     
     
         19 . The compound of  claim 16 , wherein said compound is a modulator of expression of protease C-E serine protein.  
     
     
         20 . A method of treating a patient in need of such treatment for a condition that is mediated by protease C-E, comprising administration of the compound of  claim 16 .  
     
     
         21 . A kit comprising a nucleic acid sequence selected from a group consisting of SEQ.ID.NO.:1 and SEQ.ID.NO.:2, and fragments thereof.  
     
     
         22 . A kit comprising a serine protease C-E protein having an amino acid sequence selected from the group consisting of SEQ.ID.NO.:7 and SEQ.ID.NO.:8, and fragments or derivatives thereof.  
     
     
         23 . A pharmaceutical composition comprising the protein of  claim 10 .  
     
     
         24 . The pharmaceutical composition of  claim 23  wherein said composition is a topical skin care composition.  
     
     
         25 . A non-pharmaceutical composition comprising the protein of  claim 10 .  
     
     
         26 . The non-pharmaceutical composition of  claim 25  wherein the formulation is selected from the group consisting of laundry detergent, shampoo, hard surface cleaning compositions, and dish care cleaning composition.  
     
     
         27 . A method of treating an imbalance of desquamation comprising topical application of the composition of  claim 24.