CA2616940A1

CA2616940A1 - Novel serine protease bssp2

Info

Publication number: CA2616940A1
Application number: CA002616940A
Authority: CA
Inventors: Hidetoshi Uemura; Akira Okui; Katsuya Kominami; Nozomi Yamaguchi; Shinichi Mitsui
Original assignee: Fuso Pharmaceutical Industries, Ltd.; Hidetoshi Uemura; Akira Okui; Katsuya Kominami; Nozomi Yamaguchi; Shinichi Mitsui
Current assignee: Fuso Pharmaceutical Industries Ltd
Priority date: 1998-11-20
Filing date: 1999-11-19
Publication date: 2000-06-02
Anticipated expiration: 2019-11-19
Also published as: CA2616940C

Abstract

There are provided proteins having amino acid sequences represented by SEQ ID NOS: 2, 4, 6, 8 and 10;
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 BSSP2; an antibody against BSSP2; and a method for detecting BSSP2 in a specimen by using the antibody.

Claims

1. A protein comprising an amino acid sequence of 240 amino acids represented by the 1st to 240th amino acids of SEQ ID NO: 10; or a protein comprising an amino acid sequence derived from the amino acid sequence represented by the 1st to 240th amino acids of SEQ ID NO: 10 by deletion, substitution or addition of one to several amino acids and having similar properties to that of the protein comprising the amino acid sequence represented by the 1st to 240th amino acids of SEQ ID NO: 10; or a modified derivative thereof.

2. A nucleotide sequence represented by the 807th to 1526th bases of SEQ ID NO: 9; a nucleotide sequence encoding an amino acid sequence represented by the 1st to 240th amino acids of SEQ ID NO: 10; or a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence under stringent conditions and encoding a protein having similar properties to that of a protein comprising the amino acid sequence represented by the 1st to 240th amino acids of SEQ
ID NO: 10.

3. A protein comprising an amino acid sequence of 457 amino acids represented by the -217th to 240th amino acids of SEQ ID NO: 10; or a protein comprising an amino acid sequence derived from the amino acid sequence represented by the -217th to 240th amino acids of SEQ ID
NO: 10 by deletion, substitution or addition of one to several amino acids and having similar properties to that of the protein comprising the amino acid sequence represented by the -217th to 240th amino acids of SEQ ID NO: 10; or a modified derivative thereof.

4. A nucleotide sequence represented by the 156th to 1526th bases of SEQ ID NO: 9; a nucleotide sequence encoding an amino acid sequence represented by the -217th to 240th amino acids of SEQ ID NO: 10; or a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence under stringent conditions and encoding a protein having similar properties to that of a protein comprising the amino acid sequence represented by the -217th to 240th amino acids of SEQ ID NO: 10.

5. A protein comprising an amino acid sequence of 217 amino acids represented by the -217th to -1st amino acids of SEQ ID NO: 10; or a protein comprising an amino acid sequence derived from the amino acid sequence represented by the -217th to -1st amino acids of SEQ ID
NO: 10 by deletion, substitution or addition of one to several amino acids and having similar properties to that of the protein comprising the amino acid sequence represented by the -217th to -1st amino acids of SEQ ID NO: 10; or a modified derivative thereof.

6. A nucleotide sequence represented by the 156th to 806th bases of SEQ ID NO: 9; a nucleotide sequence encoding an amino acid sequence represented by the -217th to -1st amino acids of SEQ ID NO: 10; or a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence under stringent conditions and encoding a protein having similar properties to that of a protein comprising the amino acid sequence represented by the -217th to -1st amino acids of SEQ ID NO: 10.

7. A nucleotide sequence represented by SEQ ID
NO: 9; or a nucleotide sequence hybridizable with a nucleotide sequence which is complementary to the above nucleotide sequence under incubation in a solution containing 5 × SSC, 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 and encoding a protein having similar properties to that of a protein encoded by the nucleotide sequence represented by SEQ ID NO: 9.

8. A vector comprising a promoter and a polynucleotide operably linked thereto having the nucleotide sequence described in any one of claims 2, 4, 6 and 7.

9. A cell comprising the nucleotide sequence of any one of claims 2, 4, 6 and 7 which is transformed with the vector of claim 8.

10. A process for producing a protein which comprises culturing cells transformed with the vector of claim 8, and collecting mouse Brain-specific Serine Protease 2 (mBSSP2) produced by the cells.

11. A process for producing a protein, which comprises culturing cells transformed with the nucleotide sequence described in any one of claim 2, 4, 6 and 7, and collecting human Brain-Specific Serine Protease 2 (hBSSP2) produced by the cells.

12. The process according to claim 10 or 11, wherein the cells are E. coli cells, animal cells or insect cells.

13. A cell whose expression level of BSSP2 gene has been altered, wherein the BSSP2 gene comprises SEQ ID
NO: 9.

14. The cell according to claim 13, wherein BSSP2 gene is cDNA, genomic DNA or synthetic DNA encoding BSSP2.

15. The cell according to claim 13, wherein the expression level of said BSSP2 gene has been altered by mutating a gene expression regulatory site.

16. A cell of a knockout mouse whose BSSP2 gene function is deficient, wherein the BSSP2 gene comprises SEQ
ID NO: 9.

17. An antibody against the protein described in any one of claims 1, 3 and 5.

18. The antibody according to claim 17 which is a polyclonal antibody, a monoclonal antibody or a peptide antibody.

19. A process for producing a monoclonal antibody against the protein described in any one of claims 1, 3 and 5 which comprises administering the protein according to any one of claims 1, 3 and 5 or a fragment thereof to a warm-blooded animal other than a human being, selecting the animal whose antibody titer is recognized, collecting its spleen or lymph node, fusing the antibody producing cells contained therein with myeloma cells to prepare a monoclonal antibody producing hybridoma.

20. A method for determining a presence of the protein described in any one of claims 1, 3 and 5 in a specimen, the method comprising assessing immunological binding of an antibody against the protein in the specimen.

21. A method for determining a presence of hBSSP2 or a fragment thereof in a specimen, which comprises reacting a monoclonal antibody or a polyclonal antibody against the protein described in any one of claims 1, 3 and 5 or a fragment thereof and a labeled antibody with hBSSP2 or a fragment thereof in the specimen to detect a sandwich complex thus produced.

22. A method for determining a presence of hBSSP2 or a fragment thereof in a specimen, which comprises reacting a monoclonal antibody or a polyclonal antibody against the protein described in any one of claims 1, 3 and 5 or a fragment thereof with labeled hBSSP2 or a fragment thereof in the specimen to detect an amount of hBSSP2 or a fragment thereof in the specimen, based on an amount of the labeled hBSSP2 reacted with the antibody.

23. The method according to any one of claims 20 to 22, wherein the specimen is a body fluid.

24. A diagnostic marker for diseases in tissues comprising the protein according to any one of claims 1, 3 and 5.