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Structure and functions of a very small tetraheme protein -Molecular mechanism of bioremediation-

Research Project

Project/Area Number 10044072
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionYokohama National University

Principal Investigator

AKUTSU Hideo  YNU, Faculty of Engineering, Professor, 工学部, 教授 (60029965)

Co-Investigator(Kenkyū-buntansha) OZAWA Kiyoshi  YNU, Faculty of Engineering, Research Associate, 工学部, 助手 (20251770)
FUJIWARA Toshimichi  YNU, Faculty of Engineering, Associate Professor, 工学部, 助教授 (20242381)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1998: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordscytochrome cィイD23ィエD2 / tetraheme protein / Nuclear Magnetic Resonance / redox potential / over expression system / electron transfer / electron transport protein / シトクロム / Cgf C_3 / CgbC_3 / チヘムタンパク質
Research Abstract

The most remarkable characteristic of many Shewanella species is their ability to reduce insoluble metal oxides, including iron, and manganese. S. putrefaciens strain MR1 was specifically isolated as a metal oxide (iron and manganese) reducing organism, and shown to couple the reduction of metal oxides to the oxidation of organic carbon. The ability of Shewanella species to grow with a variety of different electron acceptors having redox potentials from +0.8 V (oxygen) to about 0 (elemental sulfur) implies that this bacterium is able to adjust to growth conditions through changes in the activity and/or the composition of the electron transport chain. Cytochromes are abundant in Shewanella and have been implicated in the reduction of metal oxides. The redox behavior of a tetraheme protein, cytochrome cィイD23ィエD2 was investigated in this work. The macroscopic and microscopic redox potentials were determined at pH 9. Although they are low in comparison with other cytochromes, they are higher than the redox potentials of cytochrome cィイD23ィエD2 from Desulfovibrio vulgaris Miyazaki F (DvMF). DvMF cytochrome cィイD23ィエD2 gene was successfully expressed in a facultative aerobe, Shewanella putrefaciens MR-I, under anaerobic, microaerobic, and aerobic conditions with good yields. A derivative of the broad-host-range plasmid pRK415 containing the cytochrome cィイD23ィエD2, genes from DvMF was used for transformation of S. putrefaciens MR-1, resulting in the production of holocytochrome cィイD23ィエD2 that was indistinguishable from that produced by D. vulgaris Miyazaki F.

Report

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

    (29 results)

All Other

All Publications (29 results)

  • [Publications] S. Kim: "The interactions of ferric and ferrous cytochrome c with cardiolipin in phospholipid membranes studied by solid-state ^2H and ^<31>P NMR"J. Mol. Struct.. 441. 183-188 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ishida: "A Primitive pathway of porphyrin biosynthesis and enzymology in Desulfovibrio vulgaris"Proc. Natl. Acad. Sci. USA. 95. 4853-4858 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ohmura: "Paramagnetic Inversion of the Sign of the Interference Contribution to the Transverse Relaxation of the Imido Protons of the Coordinated Imidazoles in the Uniformly ^<15>N Labeled Cytochrome c_3"J. Magn. Reson.. 131. 367-372 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ohmura: "Ionic-strength dependent physicochemical factors in cytochrome c_3 regulating the electron transfer rate"Biophys. J.. 75. 1483-1490 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Yagi: "Functional conformational changes in the TF_1 ATPase β subunit probed by twelve"Biophys. J.. 77. 2175-2183 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Yoshida: "Molecular Mobility of Protein in Lyophilized Formulations Linked to the Molecular Mobility of Polymer Excipients, as Determined by High Resolution Solid-State ^<33>C NMR"Pharmaceutical Res.. 16. 1621-1625 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 阿久津秀雄、河野敬一: "多核NMR-自然を複眼で見る-"廣川書店. 147 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kim, K. Shin, T. Fujiwara and H. Akutsu: "The interactions of ferric and ferrous cytochrome c with cardiolipin in phospholipid membranes studied by solid-state ィイD12ィエD1H and 31ィイD1PィエD1 NMR"J. Mol. Struct.. 441. 183-188 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ishida, L. Yu, H. Akutsu, K. Ozawa, S. Kawanishi, A. Seto, T. Inubushi and S. Sano: "A primitive pathway of porphyrin biosynthesis and enzymology in Desulfovibrio vulgaris"Proc. Natl. Acad. Sci. USA. 95. 4853-4858 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ohmura, E. Harada, T. Fujiwara, G. Kawai, K. Watanabe and H. Akutsu: "Paramagnetic Inversion of the sign of the interference contribution to the transverse relaxation of the imido protons of the coordinated imidazoles in the uniformly ィイD115ィエD1N labeled cytochrome cィイD23ィエD2"J. Magn. Reson.. 131. 367-372 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Ohmura, H. Nakamura, K. Niki, M. A. Csanovich and H. Akutsu: "Ionic-strength dependent physicochemical factors in cytochrome cィイD23ィエD2 regulating the electron transfer rate"Biophys. J.. 75. 1483-1490 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] h, Yagi, K. Tozawa, N. Sekino, T. Iwabuchi, M. Yoshida, and H. Akutsu: "Functional conformational changes in the TFィイD21ィエD2ATPase βsubunit probed by twelve tyrosine residues"Biophys. J.. 77. 2175-2183 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Yoshioka, Y. Aso, S. Kojima, S. Sakurai, T. Fujiwara, and H. Akutsu: "Molecular mobility of protein in lyophilized formulations linked to the molecular mobility of polymer excipients, as determined by high resolution solid-state ィイD113ィエD1C NMR"Pharmaceutical Res.. 16. 1621-1625 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K. Ozawa, T. Meikari, K. Motohashi, M. Yoshida, and H. Akutsu: "Evidence for the presence of F-type ATPsynthase involved in sulfate-respiration in Desulfovibrio vulgais"J. Bacteriol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Akutsu and K. Kawano: "Multinuclear NMR (in Japanese)"Hirokawashoten, Tokyo. (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S.Kim: "Osmotic stability of muratnyl dipeptide-bearing liposomes and molecular miscibility in their membranes"Bull. Chem. Soc. Jpn.. 72. 541-548 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] H.Yagi: "Functional conformational changes in the TF_1 ATPase β subunit probed by twelve tyrosine residues"Biophys. J.. 77. 2175-2183 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Fukushima: "Structural analysis of mastoparan by solid-state nuclear magnetic resonance in "Peptide Science- Present and Future " (ed. Y. Shimonishi)"Kluwer Acad. Pub.. 294-296 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.Yoshioka: "Molecular mobility of protein in lyophilized formulations linked to the molecular mobility of polymer excipients, as determined by high resolution solid-state ^<13>C NMR"Pharmaceutical Res.. 16. 1621-1625 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Ozawa: "Evidence for the presence of F-type ATPsynthase involved in sulfate-respiration in Desulfovibrio vulgaris"J. Bacteriol.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] T.Fujiwara: "Compound radio-frequency driven recoupling pulse sequences for effcient magnetization transfer by homonuclear dipolar interaction under magic angle spinning conditions"J. Magn. Reson.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] 阿久津秀雄,河野敬一: "多核NMR 自然を複眼で見る"広川書店. 147 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.Kim: "The interactlons of forric and ferrous cytochrome c with cardiolipin in phosphollpid membrancs studled by solid-state ^2H and ^<31>P NMR." J.Mol.Struct.441. 183-188 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Odahara: "Membrane dynamica in the intact PM2 phage and its host colls as monitored by T_1p(H)." Biochim.Biophys.Acta. 1369. 240-246 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Ishida: "A primitlve pathway of porphyrin biosynthesis and enzymology in Desulfovibrlo vulgarls." Proc.Natl.Acad.Sci.USA. 95. 4853-4858 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Ohmura: "Paramagnetic Inversion of the sign of the interference contrlbutlon to the transvarse relaxation of the imido Protons of the eoordinated imldazoles in the uniformly ^<13>N labeled cytochrome c_3" J.Magn.Reson.131. 367-372 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Fujiwara: "Multidimensional solid-state NMR for determining dihedral angle from the correlation of ^<13>C-^1H and ^<13>C-^<13>C dipolar.interactions under magic-angle spinning conditions." J.Chem.Phys.109. 2380-2393 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Ohmura: "Ionie-strength dependent physlcochemical factors in cytochrome c_3 regulating the electron transfer rata." Biophys.J.75. 1483-1490 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 阿久津秀雄: "構造生物学とその解析法(分担)" 共立出版株式会社, 14 (1997)

    • Related Report
      1998 Annual Research Report

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

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