| Project/Area Number |
09480153
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Tokyo University of Pharmacy and Life Science |
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Principal Investigator |
TAKAHASHI Kenji Tokyo University of Pharmacy and Life Science School of Life Science Professor, 生命科学部, 教授 (70011533)
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| Co-Investigator(Kenkyū-buntansha) |
NISHII Wataru Tokyo University of Pharmacy and Life Science School of Life Science Professor, 生命科学部, 助手 (30287461)
KOJIMA Masaki Tokyo University of Pharmacy and Life Science School of Life Science Professor, 生命科学部, 助手 (90277252)
INOUE Hideshi Tokyo University of Pharmacy and Life Science School of Life Science Professor, 生命科学部, 助教授 (20184765)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Brain-gut proteinase / Progastrin-cleaving proteinase / CAAX-motif-specific proteinase / Hepsin / Colopsin / Cathepsin E / Membrane-bound proteinase / Proteinase / エンテロペプチダーゼ / カテプシンE遺伝子 / 膜結合性プロテアーゼ / セリンプロテアーゼ / CAAX-モチーフ特異的プロテアーゼ / トリプシン / α-キモトリプシン / 基質特異性 / 金属プロテアーゼ |
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| Research Abstract |
1. A novel membrane-bound mctalloendopeptidase cleaving progastrin C-terminal-(88-101) peptide as well as bradykinin and BAM-12P and an endopeptidase specific for the C-terminal CAAX motif of Ras and related proteins were solubilized from porcine and bovine brains, respectively, purified and characterized.
2. Enteropeptidase was solubilized from porcine duodenum, purified and characterized with special reference to the carbohydrate moieties ; the specificity toward various synthetic peptide substrates was sysytematically investigated.
3. The peptide bond specificities of pancreatic trypsin and alpha-chymotrypsin were investigated at high pH in order to restrict the cleavage specificity ; they were shown to selectively cleave Arg-X and Phe-X bonds, respectively, at a high alkaline pH (pH-13).
4. The microsomal membrane-bound serine proteinase from rat liver was identified as hepsin from analyses of its polypeptide chain composition and partial amino acid sequence.
5. The tissue distribution of colopsin (a novel membrane-bound serine proteinase from intestine) was investigated with human and rat tissues.
6. The gene for mouse cathepsin E was isolated and sequenced, and its chromosomal location was clarified. In addition, the substrate specificity at neutral pH and interaction with alpha 2-macroglobulin of cethepsin E from human gastric mucosa were investigated.
7. Pepsinogen isoforms were isolated and characterized from the gastric mucosae of coelacanth stomach and the N-terminal amino acid sequences were determined.
8. In relation to this project, additional characterization studies were performed on some other proteinases including nepenthesin, flarial proteinases, mold acid proteinases A and B, bacterial metalloendopeptidases and siganal peptidase and bromelain inhibitors.
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