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Systematic search for novel proteinases in gastrointestinal tract and studies on their physiological roles and structure/function

Research Project

Project/Area Number 07458149
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionTokyo University of Pharmacy and Life Science

Principal Investigator

TAKAHASHI Kenji  Tokyo University of Pharmacy and Life Science, Department of Life Science, Professor, 生命科学部, 教授 (70011533)

Co-Investigator(Kenkyū-buntansha) KOJIMA Masaki  Tokyo University of Pharmacy and Life Science, Department of Life Science, Assis, 生命科学部, 助手 (90277252)
INOUE Hideshi  Tokyo University of Pharmacy and Life Science, Department of Life Science, Assis, 生命科学部, 助教授 (20184765)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥5,400,000 (Direct Cost: ¥5,400,000)
Keywordsgastrointestinal proteinase / VIP-degrading endopeptidase / gastric trypsin / enteropeptidase / hepsin / pepsinogen / _2-macroglobulin / trypsin / プロガストリン分解プロテアーゼ / α_2マクログロブリン / プロリルエンドペプチダーゼ / αキモトリプシン / β-アミロイドペプチド / 細胞内トリプシン / プロセシング / 蛋白分解
Research Abstract

1.A novel membrane-bound serine proteinase degrading vasoactive intestinal polypeptide fairly specifically and a novel membrane-associated intracellular trypsin were isolated from porcine gastric antral mucosa and characterized.
2.Enteropeptidase cDNA was isolated and cloned from the cDNA library of rat duodenum and its primary structure was deduced. In addition, the tissue distribution
of its mRNA was analyzed.
3.A novel membrane-associated metalloproteinase degrading progastrin peptide fairly specifically was isolated from porcine brain and characterized.
4.The microsomal membrane-bound serine proteinase from rat liver was identified as hepsin from analyzes of its polypeptide chain composition and partialamino acid sequence.
5.Pepsinogen isoforms were isolated and characterized from the gastric mucosae of turtle and tuna stomach and the amino acid sequences of their major forms were determined.
6.Normal human blood alpha _2-macroglobulin was shown to contain a small but significant amount of novel serine proteinase (s) naturally entrapped in it.
7.Pancreatic trypsin and alpha-chymotrypsin were shown to selectively cleave Arg-X and Phe-X bonds, respectively, at a high alkaline pH (pH-13).
8.In relation to this project, additional characterization studies were performed on some other proteinases including follipsin, prolylendopeptidase, acid proteinase B,and signal peptidase and bromelain inhibitors.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (50 results)

All Other

All Publications (50 results)

  • [Publications] Gwang-Ho Jeohn: "Purification and characterization of a Vasoattive Intestinal Polypeptide-degrading Endoprotease from Poricine Antral Mucosal Membranes" Journal of Biological Chemistry. 270. 7809-7815 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Gwang-Ho Jeohn: "Isolation and Characterization of Gastric Trypsin from the Microsomal Fraction of Porcine Gastrid Antral Mucosa" Journal of Biological Chemistry. 270. 14748-14755 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Junji Onishi: "Cleavage Specificity of Porcine Follipsin" Journal of Biological Chemistry. 270. 19391-19394 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ken-ichi Hatano: "Prinary Structure Sequence-specific ^1H-NMR Assignments and Secondary Structure in Solution of Bromelain Inhibitor VI Pineapple Stem" European Journal of Biochemistry. 232. 335-343 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yong-Tae Kim: "Identification of Trp 300 as an Important Residue for Escherichia coli Leader Peptidase Activity" European Journal of Biochemistry. 234. 358-362 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Gwang-Ho Jeohn: "Specific Cleavage of the Gln^<15>-Lys^<16> Bond in β-Amyloid Peptidase by porcine Vasoactive Intestinal Polypeptide-degrading Endopeptidase" Protein and Peptide Letters. 3. 101-106 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Naohisa Yahagi: "Complementary DNA Cloning and Sequencing of Rat Entero peptidase and Tissue Distribution of Its mRNA" Biochemical and Biophysical Research Communications. 219. 806-812 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akira Hirasawa: "Purification and Characterization of Turfle Pepsinogen and Pepsin" Journal of Biochemistry. 120. 407-414 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Masao Tanji: "The Primary Structure of the Major Pepsinogen from the Gastric Mucosa of Tuna Stomack" Journal of Biochemistry. 120. 647-656 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yasuko Sakurai: "Trypsin-like Endopeptidasecs Naturally Entopped in Human Blood α_2-Macroglobulin" Biomedical Research. 17. 347-350 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takayuki Takahashi: "Prolye Endopeptidase from the Follicular Flaid of Porcine Ovary : Comparison with the Liver Enzyme" Biomedical Research. 17. 435-442 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Inoue: "Heterologous Expression and Site-directed Mutagenesis Studies on the Activation Mechanism and the Roles of the Basic Residues in the Prosegment of Aspergillopepsinoger I" European Journal of Biochemistry. 237. 719-725 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ken-ichi Hatano: "Solution Structure of Bromelain Inhibitor VI Pineapple Stem : Structural Similarity with Bowman-Birk Trypsin/Chymotrypsin Inhibitor from Soybeaen" Biochemistry. 35. 5379-5384 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ken-ichi Hatano: "Tertiary Structure Comparison of Bromelain Inhibitor VI from Pineapple Sten with Bovine Pancreatic Trypsin Inhibitor" Proceedings of the Japan Academy. 72. 104-107 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hee-Jung Kim: "Purification and Characterization of an Extracellular Metalloprotease from Pseudomonas fluorescens" Journal of Biochemistry. 121. 82-88 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kenji Takahashi: "The Specificity of Peptide Bond Cleavage of Acid Proteinase A from Aspergillus niger var.macrosporus toward Oxidiged Ribonuclease A" Bioscience,Biotechnology and Biochemistry. 61. 381-383 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hidemi Matsuzaki: "Evidence for the Identity of Rat Liver Microsomal Membrance-bound Serine Proteinase with Hepsin" Protein and Peptide Letters. 4(in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kenji Takahashi: "Restriction of the Specifications of Trypsin and α-Chynofrypsin at a High Alkaline pH Values" Protein and Peptide Letters. 4(in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Gwang-Ho Jcohn: "Purification and Characterization of a Vasoactive Intestinal Polypeptide-Degrading Endoprotease from Porcine Antral Mucosal Membranes" J.Biol.Chem.270. 7809-7815 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Gwang-Ho Jcohn: "Isolation and Characterization of a Gastric Trypsin from the Microsomal Fraction of Porcine Antral Mucosa" J.Biol.Chem.270. 14748-14755 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Junji Ohnishi: "Cleavage Specificity of Porcine Follipsin" J.Biol.Chem.270. 19391-19394 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ken-ichi Hatano: "Primary Structure, Sequence-Specific ^1H-NMR Assignments and Secondary Structure in Solution of Bromelain Inhibitor VI from Pineapple Stem" Eur.J.Biochem.232. 335-343 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yong-Tac Kim: "Identification of Trp300 as an Important Residue for Escherichia coli Leader Peptidase Activity" Eur.J.Biochem.234. 358-362 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Gwan-Ho Jeohn: "Specific Cleavage of the Gln^<15>-Lys^<16> Bond in beta-Amyloid Peptides by Porcine Vasoactive Intestinal Polypeptide-degrading Endopeptidase" Protein and Peptide Lett.3. 101-106 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Naohisa Yahagi: "Complementary DNA Cloning and Sequencing of Rat Enteropeptidase and Tissue Distribution of Its mRNA" Biochem.Biophys.Res.Commun.219. 806-812 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akira Hirasawa: "Purification and Characterization of Turtle Pepsinogen and Pepsin" J.Biochem.120. 407-414 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Masao Tanji: "The Primary Structure of the Major Pepsinogen from the Gastric Mucosa of Tuna Sromach." J.Biochem.120. 647-656 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yasuko Sakurai: "Trypsin-like Endopeptidase (s) Naturally Entrapped in Human Blood 2-Macroglobulin" Biomed.Res.17. 347-350 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takayuki Takahashi: "Prolyl Endopeptidase from the Follicular Fluid of Porcine Ovary : Comparison with the Liver Enzyme" Biomed.Res.17. 435-442 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Inoue: "Heterologous Expression and Site-Directed Mutagensis Studies on the Activation Mechanism and the Roles of the Basic Residues in the Prosegment of Aspergillopepsinogen I" Eur.J.Biochem.237. 719-725 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ken-ichi Hatano: "Solution Structure of Bromelain Inhibitor VI from Pineapple Stem : Structural Similarity with Bowman-Birk Trypsin/Chymotrypsin Inhibitor from Soybean" Biochemistry. 35. 5379-5384 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ken-ichi Hatano: "Tertiary Structure Comparison of Bromelain Inhibitor VI from Pineapple Stem with Bovine Pancreatic Trypsin Inhibitor" Proc.Japan Acad.72. 104-107 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hee-Jung Kim: "Purification and Characterization of an Extracellular Metallo-protease from Pseudomonas fluorescens" J.Biochem.121. 82-88 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kenji Takahashi: "The Specificity of Peptide Bond Cleavage of Acid Proteinase A from Aspergillus var.macrosporus toward Oxidized Ribonuclease A" Biosci.Biotech.Biochem.61. 381-383 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hidemi Matsuzaki: "Evidence for the Identity of the microsomal membrane-bound serine proteinase from rat liver as hepsin" Protein and Peptide Lett.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kenji Takahashi: "Restriction of the Specificities of Trypsin and alpha-Chymotrypsin at a High Alkaline pH Value" Protein and Peptide Lett.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akira Hirasawa: "Purification and characterizetion of Turtle Pepsinogen and Pepsin" Journal of Biochemistry. 120. 407-414 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Masao Tanji: "The Primary Strucfue of the Major Pepsinogen from the Gastric Mucosa of Tuna Stomach" Journal of Biochemistry. 120. 647-656 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yasuko Sakurai: "Frypsin-Like Endopeptidase(s) Naturally Entrapped in Human Blood α_2-Macroglobulin" Biomedical Peseanch. 17. 347-350 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takayuki Takahashi: "Prolyl endopeptidase from the Follieular Fluid of Porcine Ovary Comparison with the Liver Enzyme" Biomedical Pesearch. 17. 435-442 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hidemi Matsuzaki: "Evidence for the Identity of Rat Liver Microsomal Membrane-Bound Serine Proteinase with Hepsin" Protein and Peptide Letters. 4(in prees). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kenji Takahashi: "Pestriction of the Specifi cities of Trypsin and α-Chymotrgpsin at a High Alkaline pH Values" Protein and Peptide Letters. 4(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Gwang-Ho Jeohn: "Purification and Characterization of a Vasaactive Intestinal Polypeptide-degrading Endoprotease from Porcine Antral Mucosal Membranes" Journal of Biological Chemistry. 270. 7809-7815 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Gwang-Ho Jeohn: "Isolation and Characterization of Gastric Trypsin from the Microsomal Fraction of Porcine Gastric Antral Mucosa" Journal of Biological Chemistry. 270. 14748-14755 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Junji Ohnishi: "Cleavage Specificity of Porcine Follipsin" Journal of Biological Chemistry. 270. 19391-19394 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kenji Takahashi: "Proteinase A from Aspergillus niger" Mothods in Enzymology. 248. 146-155 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yong-Tae Kim: "Leader Peptidase from Escherichia coli: Overexpression, Characterization, and Inactivation by Modification of Fryphphan Residues 300 and 310 with N-Bromos uccinimide" Journal of Biochemistry. 117. 535-544 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yong-Tae Kim: "Identification of Trp300 as an important residwe for Escherichia coli leader peptidase activity" European Journal Biochemisty. 234. 358-362 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Gwang-Ho Jeohn: "Specific Cleavage of the Gln^<15>-Lys^<16>Bond in β-Amyloid Peptides by Porcine Vasoactive Intestinal Polypeptide-degrading Endopeptidase" Protein and Peptide Letters. 1(in press). (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Naohisa Yahagi: "Complementary DNA Cloning and Seguancing of Rat Enteropeptidase and Tissue Distribution of Its mRNA" Biochemical and Biophysical Research Communication. (in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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