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Structural Study on Electron Tunneling and Proton Relay in Proteins by Using Ultraviolet Resonance Raman Spectroscopy

Research Project

Project/Area Number 60470018
Research Category

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

Allocation TypeSingle-year Grants
Research Field 構造化学
Research InstitutionOkazaki National Research Institutes

Principal Investigator

KITAGAWA Teizo  Institute for Moleculr Science, その他, 教授 (40029955)

Co-Investigator(Kenkyū-buntansha) KAMOGAWA Keiji  Institue for Molecular Science, 分子科学研究所, 助手 (40150057)
Project Period (FY) 1985 – 1986
Project Status Completed (Fiscal Year 1986)
Budget Amount *help
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1986: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1985: ¥6,000,000 (Direct Cost: ¥6,000,000)
KeywordsResonance Raman / Bacteriorhodopsin / Cytochrome oxidase / Proton pump / Electron transfer / 電子伝達
Research Abstract

Molecular mechanism of the vectorial proton transportation through biological membrane was investigated by using resonance Raman spectroscopy, and a system for measuring UV resonance Raman spectra of protein moieties was constructed. Two typical systems were picked up; one is a light driven proton pumping system having bacteriorhodopsin and the other is an electron-driven proton pumping system containing cytochrome c oxidase. For bacteriorhodopsin intermediates involved in a time region when the Schiff base proton of the chromophore was transferred to the protein moiety were examined and discussion was focused on what would happen when the protein moiety did not accept the proton. For cytochrome c oxidase the occurrence of photoreduction was confirmed and the electron transfer rate from the a heme to the <a_3> heme was determined. It was found that the aerobic photoreduction generates a photosteady state composed mainly of <a^(2+)> <a(_3^(3+))> , and that its <a^(2+)> and <a(_3^(3+))> hemes can be selectively probed with the 441.6 and 406.7 nm excitations, respectively. Sensitivity of the Fe-His stretching Raman intensity to the enzymic activity was pointed out, and transient Raman spectra of oxygenated intermediates were discussed. A system to observe Raman spectra excited by UV light around 200-240 nm was set up with a SPEX 1260 single monochromator equipped with a 1200 gr/mm and 200 nm blaze grating. Excitation light was derived from Nd:YAG laser and Raman shifter. The system brought about spectra of good quality for aromatic amino acid residues.

Report

(1 results)
  • 1986 Final Research Report Summary
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] A.Maeda;T.Ogurusu;T.Yoshizawa;T.Kitagawa: Biochemistry. 24. 2517-2521 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Ogura;S.Yoshikawa;T.Kitagawa: Biochemistry. 24. 7746-7752 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Ogura;S.Yoshikawa;T.Kitagawa: Biochimica Biophysica Acta. 832. 220-223 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] N.Sone;T.Ogura;T.Kitagawa: Biochimica Biophysica Acta. 850. 139-145 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] S.Hashimoto;Y.Tatsuno;T.Kitagawa: Proceedings of National Academy of Sciences of U.S.A.83. 2417-2421 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] A.Maeda;T.Ogura;T.Kitagawa: Biochemistry. 25. 2798-2803 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Kitagawa: "Spectroscopy of Biological Systems edited by R.J.H.Clark and R.E.Hester" Wiley and Sons, (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Kitagawa: "Biomolecules-Electronic Aspects edited by C.Nagata,M.Hatano,J.Tanaka and H.Suzuki." Japan Scientific Society Press, (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] A. Maeda, T. Ogurusu, T. Yoshizawa, and T. Kitagawa: "Resonance Raman Study on Binding of Chloride to the Chromophore of Halorhodopsin" Biochemistry. 24. 2517-2521 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T. Ogura, S. Yoshikawa, and T. Kitagawa: "Resonance Raman Study on Photoreduction of Cytochrome c Oxidases; Distinction of the a and <a_3> Hemes." Biochemistry. 24. 7746-7752 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T. Ogura, S. Yoshikawa, and T. Kitagawa: "Resonance Raman Spectra of Oxygenated Intermediates of Cytochrome c Oxidase Detected with a Mixed Flow Transient Raman Apparatus." Biochimica et Biophysica Acta. 832. 220-223 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] N. Sone, T. Ogura, and T. Kitagawa: "Iron-Histidine Stretching Raman Line and Enzymic Activities of Beef-and Bacterium Cytochrome c Oxidase" Biochimica et Biophysica Acta. 850. 139-145 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] A. Maeda, T. Ogura, and T. Kitagawa: "Resonance Raman Study on Proton-Dissociated State of Bacteriorhodopsin: Stabilization of L-like Intermediate Having the all-trans Chromophore." Biochemistry. 25. 2798-2803 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary

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Published: 1987-03-31   Modified: 2016-04-21  

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