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2001 Fiscal Year Final Research Report Summary

Studies on Multi-nuclear Copper Enzymes and their Model Complexes

Research Project

Project/Area Number 11440198
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Inorganic chemistry
Research InstitutionOsaka University

Principal Investigator

SUZUKI Shinnichiro  Osaka University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (70116052)

Co-Investigator(Kenkyū-buntansha) KATAOKA Kunishige  Kanazawa University, Faculty of Science, Assistant Professor, 理学部, 講師 (40252712)
YAMAGUCHI Kazuya  Osaka University, Graduate School of Science, Assistant Professor, 大学院・理学研究科, 講師 (80252550)
Project Period (FY) 1999 – 2001
KeywordsMulti-nuclear Copper Enzymes / denitrifying bacteria / nitrite reductase / type 1 Cu / type 2 Cu / pulse radiolysis / nitrous oxide reductase / intramolecular electron transfer process
Research Abstract

The purpose of this project is to clarify the structure and function of Cu-containing nitrite reductase and nitrous oxide reductase.
1) In order to shed light on the His255 and Asp98 residues around the type 2 Cu center in blue Achromobacter xylosoxidans nitrite reductase, two mutants were expressed in E. coil.
The Asp98Ala and His255Ala mutants show the very low enzyme activities compared with the wild enzyme at pH 5.5, suggesting that these residues play a role of proton donation for the formation of water molecule.
The Asp and His255 residues are also very important for the control of the intramolecular electron transfer rate from type 1 Cu to type 2.
2) The nitrite reductase from Hyphomicrobium sp. A3151 has the two type 1 Cu's and one type 2 Cu, which is different from the other nitrite reductases containing type 1 Cu : type 2 Cu = 1 : 1. One of type 1 Cu(II) ions is blue and the other is green.
3) The nitrous oxide reductase from Hyphomicrobium sp. A3151 is aerobically stable, and there are two forms, purple oxidized and blue reduced forms.
4) The type 2 Cu(I) and (II) model complexes were prepared. The Cu(I) complex binding nitrite with nitrogen atom stoichiometrically gives NO gas in the presence of proton.

  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Deligeer, K.Kataoka, K.Yamaguchi, S.suzuki: "Spectroscopic and Electrochemical Properties of Cytochrome c551 from Alcaligenes sylosoxidans GIFU 1051"Bulletin of the Chemical Society of Japan. 73. 1839-1840 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Kataoka, A.Kondo, K.Yamaguchi, S.Suzuki: "Spectroscopic and Electrochemical Properties of the Met86Gln Mutant of Achromobacer cycloclastes Pseudoazurin"Journal of Inorganic Biochemistry. 82. 79-84 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Deligeer, Kunishige Kataoka, Shinnichiro Szuki: "Characterization of Azidc-Binding Type 2 Cu(11) Site of Nitrite Reductase"Bulletin of the Chemical Society of Japan. 74・1. 183-184 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Deligeer, Ryoichi fukunaga, Kunishige Kataoka, kazuya Yamaguchi, Sinnichiro Suzuki: "Spectroscopic and functional Characterization of Cu-containing netucte reductase from Hyphonic-****** A3151"Journal of Inorganic Biochemistry. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 岩本, 萩野, 久司, 山内編, 山口和也: "「大学院錯体化学」:金属酵素の構造と機能"講談社サイエンティク,東京. 283-290 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 岩本, 萩野, 久司, 山内編, 鈴木晋一郎: "「大学院錯体化学」:核酸と金属イオン、金属錯体との相互作用"講談社サイエンティク、東京. 291-296 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 吉田, 茂木編, 片岡邦重, 鈴木晋一郎: "「生体膜のエネルギー装置」:嫌気呼吸系の膜エネルギー装置"日本生化学会編,共立出版,東京. 60 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsuyoshi Inoue et al.: "Crystallization and prelininary X-ray stydy of iso-2 azurin from the methylotrophic bacterium, Methylomonas J"Acta Cryst.. D55. 307-309 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shinnichiro Suzuki et al.: "Structure-function relationships of copper-containing nitrite reductases"Coord. Chem. Rev.. 190-192. 245-265 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsuyoshi Inoue et al.: "Crystal structure determinations of oxidized and reduced pseudoazurins from Achromobacter cycloclastes : Concerted movement of copper site in redox forms with therearrangement of hydrogen bond at a remote histidine"J. Biol. Chem.. 274. 17845-17852 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuo Kobayashi et al.: "The pH-dependent changes of intramolecular electron transfer on copper-containing nitrite reductase"J. Biochem.. 126. 408-412 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shinnichiro Suzuki et al.: "Spectroscopic and electrochemical properties of two azurins (Az-iso1 and Az-iso2) from the obligate methylotroph Methylomonas sp strain J and the structure of novel Az-iso2"J. Biol. Inorg. Chem.. 4. 749-758 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kunishige Kataoka et al.: "Functional analysis of conserved asparate and histidine residues located around the type 2 Cu site of copper-containing nitrite reductase"J. Biochem.. 127. 345-350 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shinnichiro Suzuki et al.: "Intramolecular electron-transfer process of native and mutant forms of blue copper-containing nitrite reductase from Alcaligenes xylosoxidans"Inorg. Reac. Mech.. 2. 129-135 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Deligeer et al.: "Spectroscopic and electrochemical properties of cytochrome c_<551> from Alcaligenes xylosoxidans GIFU 1051"Bull. Chem. Soc. Jpn.. 73. 1839-1840 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kunishige Kataoka et al.: "Spectroscopic and electrochemical of the Met86Gln mutant of Achromobacter cycloclastes pseudoazurin"J. Inorg. Biochem.. 82. 79-84 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shinnichiro Suzuki et al.: "Metal coordination and mechanism of multi-copper nitrite reductase"Acc. Chem. Res.. 33. 728-735 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Deligeer et al.: "Characterization of azide-binding type 2 Cu(II) site of nitrite reductase"Bull. Chem. Soc. Jpn.. 74. 183-184 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Deligeer et al.: "Spectroscopic and functional characterization of Cu-containing nitrite reductase from Hyphomicrobium denitrificans A3151"J. Inorg. Biochem.. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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