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Structural Regulation Mechanism for Reactivity in Metalloproteins

Research Project

Project/Area Number 07309006
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section総合
Research Field 広領域
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

MORISHIMA Isao  Kyoto University, Graduate School of Engineering, Professor, 工学研究科, 教授 (50026093)

Co-Investigator(Kenkyū-buntansha) MOGI Tateshi  Tokyo University, Graduate School of Science, Research Associate, 理学部, 助手 (90219965)
HORI Hiroshi  Osaka University, Faculty of Engineering Science, Research Associate, 基礎工学部, 助手 (20127294)
FUKUYAMA Keiichi  Osaka University, Graduate School of Science, Professor, 理学部, 教授 (80032283)
HASE Toshiharu  Osaka University, Protein Research Institute, Professor, 蛋白質研究所, 教授 (00127276)
KITAGAWA Teizo  Institute of Molecular Science, Professor, 教授 (40029955)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥11,300,000 (Direct Cost: ¥11,300,000)
Fiscal Year 1996: ¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1995: ¥6,000,000 (Direct Cost: ¥6,000,000)
KeywordsPeroxidase / Ferredoxin / Nitrite Reductase / Oxgen Snsor Fix L / Cytochrome c Oxidase / Cytochrome P450 / 酸素センサーFixL / チトクロム酸化酵素
Research Abstract

The Primary results in this research project are as follows :
1)The extensive mutation in horseradish peroxidase has revealed that the highly conserved hydrogen network in the distal site plays a key role in catalytic activity in peroxidases. The detailed molecular mechanism in peroxidase reaction was discussed.
2)The crystal structure of ARP,which shows specific high reactivity for luminol, was resolved and the substrate binding site was proposed. By using time-resolved resonance Raman spectroscopy, we examined the intermediate species in the reaction ARP and compared them with those of horseradish peroxidase.
3)We followed the reaction of cytochrome c oxidase with hydrogen peroxide and idetified some key inter mediates in the reaction.
4)The Regulation mechanism for the terminal oxidase in Escherichia coli was investigated by using the mutants.
5)The electron transfer in nitrite reductase, cytochrome cd_1, was examined by pulse radiolysis and reduction mechanism for the enzyme was proposed.
6)The systematic mutation in ferredoxin showed the specific interaction sites between ferredoxin and sulfite reductase.
7)We resolved crystal structure of another nitrite reductase, P450nor, and discussed the molecular mechanism for reduction of nitrite by the enzyme.
8)EPR was utilized for the investigation of the interaction between cytochrome P450cam and its electron donor, putidaredoxin. Some specific interactions were elucidated.

Report

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

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Matsui, T: "Preparation and Reaction of Myoglobin Mutants Bearing Both Proximal Cysteine Ligand and Hydrophobic Distal Cavity:Protein Models for the Active Site of P-450" Biochemistry. 35. 13118-13124 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nagano, S: "The Catalytic Roles of the Distal Site Asparagine-Histidine Couple in Peroxidases" Biochemistry. 35. 14251-14258 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tanaka, M: "The Distal Glutamic Acid As Acid-Base Catalyst in the Distal Site of Horseradish Peroxidase" Biochem.Biophys.Res.Commun. 227. 393-399 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Mukai, M.: "Effects of Concerted Hydrogen Bonding of Distal Histidine on Active Site Structure of Horseradish Peroxidase;Resonance Raman Studies with Asn-70 Mutants" J.Am.Chem.Soc. 119. 1758-1766 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tanaka, M.: "Hydrogen Bond Network in the Distal Site of Peroxidases:Spectroscopic Properties of Asn 70→Asp Horseradish Peroxidase Mutant" Biochemistry. 36. 9791-9798 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tanaka, M.: "Catalytic Activities and Structural Properties of Horseradish Peroxidase Distal His42→Glu or Gln Mutant" Biochemistry. 36. 9889-9898 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsui, T., Nagano, S., Ishimori, K., et al.: "Preparation and Reaction of Myoglobin Mutants Bearing Both Proximal Cysteine Ligand and Hydrophobic Distal Cavity : Protein Models for the Active Site of P-450" Biochemistry. 35. 13118-13124 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nagano, S., Tanaka, M., Ishimori, K., et al.: "The Catalytic Roles of the Distal Site Asparagine-Histindine Couple in Peroxidases" Biochemistry. 35. 14251-14258 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tanaka, M., Ishimori, K., Morishima, I.: "The Distal Glutamic Acid As Acid-Base Catalyst in the Distal Site of Horseradish Peroxidase" Biochem.Biophys.Res.Commun.227. 393-399 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Mukai, M., Nagano, S., Tanaka, M., Ishimori, K., et al.: "Effects of Concerted Hydrogen Bonding of Distal Histidine on Active Site Structure of Horseradish Peroxidase ; Resonance Raman Studies with Asn-70 Mutants" J.Am.Chem.Soc.119. 1758-1766 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tanaka, M., Ishimori, K., Morishima, I.: "Hydrogen Bond Network in the Distal Site of Peroxidases : Spectroscopic Properties of Asn 70 * Asp Horseradish Peroxidase Mutant" Biochemistry. 36. 9791-9798 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tanaka, T., Ishimori, K., Morishima, I.Mukai, M., Kitagawa, T.: "Catalytic Activities and Structural Properties of Horseradish Peroxidase Distal His42 * Glu or Gln Mutant" Biochemistry. 36. 9889-9898 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsui,T: "Preparation and Reaction of Myoglobin Mutants Bearing Both Proximal Cysteine Ligand and Hydrophobic Distal Cavity : Protein Models for the Active Site of P-450" Biochemistry. 35. 13118-13124 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagano,S: "The Catalytic Roles of the Distal Site Asparagine-Histidine Couple in Peroxidases" Biochemistry. 35. 14251-14258 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Tanaka,M: "The Distal Glutamic Acid As Acid-Base Catalyst in the Distal Site of Horseradish Peroxidase" Biochem.Biophys.Res.Commun. 227. 393-399 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Mukai,M.: "Effects of Concerted Hydrogen Bonding of Distal Histidine on Active Site Structure of Horseradish Peroxidase ; Resonance Raman Studies with Asn-70 Mutants" J.Am.Chem.Soc. 119(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Morishima, I: "NMR Studies on Recombinant Cytochrome P450cam Mutants" Biochimie. (印刷中). (1996)

    • Related Report
      1995 Annual Research Report

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

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