Novel serine protease BSSP4

Abstract

There are provided proteins having the amino acid sequences represented by SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20; proteins having amino acid sequences derived from these amino acid sequences by deletion, substitution or addition of one to several amino acids; and nucleotide sequences encoding the same; transgenic non-human animals with altered expression level of a serine protease BSSP4; an antibody against BSSP4; and a method for detecting BSSP4 in a specimen by using the antibody.

1 . A protein selected from the group consisting of: 
 (a) a protein having the amino acid sequence composed of 270 amino acids represented by the 1 st  to 270 th  amino acids of SEQ ID NO:4;    (b) a protein having the amino acid sequence composed of 319 amino acids represented by the −49 th  to 270 th  amino acids of SEQ ID NO:4; and    (c) a protein having the amino acid sequence composed of 285 amino acids represented by the −15 th  to 270 th  amino acids of SEQ ID NO:4.    
     
     
         2 . A nucleotide sequence selected from the group consisting of: 
 (i) a nucleotide sequence represented by the 151 st  to 960 th  bases of SEQ ID NO:3;    (ii) a nucleotide sequence encoding the amino acid sequence represented by the 1 st  to 270 th  amino acids of SEQ ID NO:4;    (III) a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence (i) or (ii) under stringent conditions of incubation in a solution containing 5×SSC or SSPE, 5% Denhardt's solution (01% BSA, 0.1% Ficol 1400, 0.1% PVP), 0.5% SDS and 20 μg/ml denatured salmon sperm DNA 37° C. overnight, followed by washing with 2×SSC or SSPE containing 0.1% SDS at room temperature, and encoding a protein having the same serine protease property as that of the protein having the amino acid sequence represented by the 1 st  to 270 th  amino acids of SEQ ID NO:4;    (iv) a nucleotide sequence represented by the 4 th  to 960 th  bases of SEQ ID NO:3;    (v) a nucleotide sequence encoding the amino acid sequence represented by the −49 th  to 270 th  amino acids of SEQ ID NO:4;    (vi) a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence (iv) or (v) under stringent cnditoins of incubation in a solution containing 5×SSC or SSPE, 5% Denhardt's solution (0.1% BSA, 0.1% Ficol 1400, 0.1% PVP), 0.5% SDS and 20 μg/ml denatured salmon sperm DNA at 37° C. overnight, followed by washing with 2×SSC or SSPE containing 0.1% SDS at room temperature, and encoding a protein having the same serine protease property as that of the preotein having the amino acid sequence represented by the −49 th  to 270 th  amino acids of SEQ IDN O:4;    (vii) a nucleotide sequence represented by the 106 th  to 960 th  bases of SEQ ID NO:3;    (viii) a nucleotide sequence encoding the amino acid sequence represented by the −15 th  to 270 th  amino acids of SEQ ID NO:4;    (ix) a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence (vii) or (viii) under stringent conditions of incubation in a solution containing 5×SSC or SSPE, 5% Denhardt's solution (0.1% BSA, 0.1% Ficol 1400, 0.1% PVP), 0.5% SDS and 20 μg/ml denatured salmon sperm DNA at 37° C. overnight, followed by washing with 2×SSC or SSPE containing 0.1% SDS at room temperature, and encoding a protein having the same serine protease property as that of the protein encoded by the nucleotide sequence represented by SEQ ID NO:3.    
     
     
         3 . A vector, comprising the nucleotide sequence according to  claim 2 .  
     
     
         4 . Transformed cells having the nucleotide sequence according to  claim 2  in an expressible state.  
     
     
         5 . A process for producing a protein which comprises culturing cells transformed with the nucleotide sequence according to (1) to (xi) of  claim 2  or a fragment thereof, and collecting hBSSP4 produced.  
     
     
         6 . The process according to  claim 5 , wherein the cells are  E. coli  cells, animal cells or insect cells.  
     
     
         7 . A method for screening for an inhibitor of serine protease, comprising comparing the enzyme activity of the protein according to  claim 1  upon bringing it into contact with a candidate compound with the enzyme activity of the protein without contact with the candidate compound.  
     
     
         8 . A pharmaceutical composition, comprising the protein according to  claim 1.