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Studies on molecular mechanism of glutathione one salvage pathway of Escherichia coli

Research Project

Project/Area Number 10660083
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

SUZUKI Hideyuki  Kyoto University, Graduate School of Biostudies, Associate Professor, 生命科学研究科, 助教授 (10202136)

Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywordsactive center / Escherichia coli / Ntn-hydrolase / glutathione / cysteinylglycinase / γ-glutamyltranspeptidase / aminopeptidase / ペプチダーゼ / ジペプチド / ロイシンアミノペプチダーゼ / システイン
Research Abstract

1. We found that aminopeptidases A, B and N, and dipeptidase D with broad substrate specificity are the four cysteinylglycinases of Escherichia coli K-12 and there is no peptidase specific for the cleavage of cysteinylglycine.
2. Aminopeptidase B was purified to electrophoretic homogeneity and its enzymatic characteristics were determined. The data indicates that aminopeptidase B is a metallopeptidase. Aminopeptidase is a metallopeptidase. The activity of aminopeptidase B, which was saturated with one of above divalent cations, was enhanced on the addition of a very small amount of a second divalent cation. Cysteinylglycine was the best substrates among those we tested.
3. We identified that the catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by a novel affinity labeling agent. After the modification of the enzyme, it was separated into the large subunit and the small subunit followed by the fragmentation by lysyl endopeptidase. The fragments were analyzed by MS-MS and it was found that is the Thr-residue at the N-terminal of the small subunit is the active center of this enzyme. This result strongly suggests that γ-glutamyltranspeptidase is a new member of the N-terminal nucleophile hydrolase family.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Hideyuki Suzuki et al.: "Glutathione metabolism in Escherichia coli."J.Mol.Catal.B. 6(3). 175-184 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hideyuki Suzuki et al.: "Identification of catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by γ-monofluorophosphono derivative of glutamic acid : N-terminal Thr-391 in small subunit is the nucleophile."Biochemistry. 39(26). 7764-7771 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hideyuki Suzuki et al.: "Aminopeptidases A, B and N, and dipeptidase D are the four cysteinylglycinases of Escherichia coli K-12."Journal of Bacteriology. 183(4). 1489-1490 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 鈴木秀之,熊谷英彦: "γ-グルタミルトランスペプチダーゼ:Ntn-ハイドロラーゼスーパーファミリーの新しいメンバー."蛋白質核酸酵素. (印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hideyuki Suzuki et al.: "Purification and Characterization of Aminopeptidase B from Escherichia coli K-12."Biosci.Biotechnol.Biochem.,. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Suzuki, W.Hashimoto, and H.Kumagai: "Glutathione metabolism in Escherichia coli."J.Mol.Catal.B. 6 (3). 175-184 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Inoue, J.Hiratake, H.Suzuki, H.Kumagai, and K.Sakata: "Identification of catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by γ-monofluorophosphono derivative of glutamic acid : N-terminal Thr-391 in small subunit is the nucleophile."Biochemistry. 39 (26). 7764-7771 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Suzuki, S.Kamatani, E.-S.Kim, and H.Kumagai: "Aminopeptidases A, B and N, and dipeptidase D are the four cysteinylglycinases of Escherichia coli K-12."J.Bacteriol. 183 (4). 1489-1490 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Suzuki, and H.Kumagai: "γ-Glutamyltranspeptidase : A new member of Ntn hydrolase superfamily."Protein, Nuleic Acid and Enzyme. in press. (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Suzuki, S.Kamatani, and H.Kumagai: "Purification and Characterization of Aminopeptidase B from Escherichia coli K-12."Biosci.Biotechnol.Biochem.. in press. (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hideyuki Suzuki et al.: "Identification of catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by γ-monofluorophosphono derivative of glutamic acid : N-terminal Thr-391 in small subunit is the nucleophile."Biochemistry. 39(26). 7764-7771 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hideyuki Suzuki et al.: "Aminopeptidases A, B and N, and dipeptidase D are the four cysteinylglycinases of Escherichia coli K-12."Journal of Bacteriology. 183(4). 1489-1490 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hideyuki Suzuki: "Glutathione metabolism in Escherichia coli"Journal of Molecular Catalysis : Enzymatic B. 235. 175-184 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Hideyuki Suzuki: "Glutathione metabolism in Escherichia coli" Journal of Molecular Catalysis : Enzymatic B. 235(in press). (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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