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Preparation of single crystal of phytochrome and its preliminary structural analysis by X-ray crystallography

Research Project

Project/Area Number 02454560
Research Category

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

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionOkazaki National Research Institutes, National Institute for Basic Biology

Principal Investigator

TOKUTOMI Satoru  National Institute for Basic Biology, Department of Biological Regulation Research associate, 基礎生物学研究所, 助手 (90142009)

Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥3,800,000 (Direct Cost: ¥3,800,000)
KeywordsPhytochrome / Chromophore Structure / Chromophore orientation / Single crystal / 結晶解析
Research Abstract

Phytochrome is a chromoprotein in green plants, which serves as a photoreceptor for a variety of morphogenetic responses. In order to elucidate its three-dimensional structure. crystallographic study has been started. First, conditions for crystallization of large (114kDa, exist in a dimer) phytochrome was investigated systematically. Partly crystallized blocks (0.5mm) were obtained using ammonium sulfate as precipitant, however, preparation of large single crystals has not been successful yet. In stead, crystallization of small (59kDa, chromophoric domain) phytochrome was started, which seems promissing judging from the results obtained so far, and is to be continued.
In parallel to the above research, molecular structure of the chromophore was studied using resonance Raman scattering (Tokutomi et al., 1990, FEBS Lett., Mizutani et al., Bioahem., 1991), which showed a different protoated structure between the red- and far redabsorbing forms. A model for molecular struture of the chromophore was constructed using these new informations (Toktutomi et al., 1992, Photochem. Photobiol.), indicated elongated molecular structure. Its orientation in the dimeric phytochrome molecule, proposed based on small-angle X-ray scattering (Nakasako et al., 1990, Photochem. photobiol.), was determined based on linear dichroism of oriented molecules (Toktutomiet al., i992, Photoahem. Photobiol.), which shows that the chromophore is well contained in a flat chromophoric domain of the dimeric molecule. These results can supply useful informations to the crystallographic anlysis of phytochrome.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Nakasako,M.et al.: "Quarternary structure of pea phytochrome I dimer studied with samll-angle X-ray scattering and rotary-shadowing electron microscopy." Potochem.Photobiol.52. 3-12 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tokutomi,S.et al.: "Resonance Raman scattering of large pea phytochrome at ambient temperture.Difference in chromophore protonatiion between red-and far red-absorbing forms." FEBS Letter. 269. 341-344 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Mizutani,Y.et al.: "Resonance Raman study on intact pea phytochrome and model compounds:Evidence for proton migration during phototransformation." Biochemistry. 30. 10693-10700 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tokutomi,S.et al.: "A model for molecular structure and orientation of the chromophore in a dimenic phytochrome molecule." Photochem.Photobiol.(1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 徳富 哲: "植物の光情報受容蛋白質フィトクロムの分子構造" 組織培養. 17. 425-430 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nakasako, M. et al.: "Quarternary structure of pea phytochrome I dimer studied with samll-angle X-ray scattering and rotary-shadowing electron microscopy." Potochem. Photobiol.52. 2-12 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tokutomi, S. et al.: "Resonance Raman scattering of large pea phytochrome at ambient temperture. Difference in chromophore protonatiion between red- and far red-absorbing forms." FEBS Letter. 269. 341-344 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Mizutani, Y. et al.: "Resonance Raman study on intact pea phytochrome and model compounds : Evidence for proton migration during phototransformation." Biochemistry. 30. 10693-10700 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tokutomi S.: "Molecular Structure of phytochrome : A photoreceptor in plants." The Tissue Culture (The New Science Co. Tokyo). 17. 425-430 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tokutomi S. et al.: "A model for molecular structure and orientation of the chromophore in a dimeric phytochrome molecule." Photochem. Photobiol.(1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nakasako,M.et al.: "Quarternary structure of pea phytochrome I dimer studied with samll-angle X-ray scattering and rotary-shadowing electron microscopy." Potochem.Photobiol.52. 3-12 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] tokutomi,S.et al.: "Resonance Raman scattering of large pea phytochrome at ambient temperture.Difference in chromophore protonatiion between red- and far red-absorbing forms." FEBS Letter. 269. 341-344 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mizutani,Y.et al.: "Resonance Raman study on intact pea phytochrome and model compounds:;Evidence for proton migration during phototransformation." Biochemistry. 30. 10693-10700 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Tokutomi,S.et al.: "A model for molecular structure and orientation of the chromo-phore in a dime〓ic phytochrome molecule." Photochem.Photobiol.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 徳富 哲: "植物の光情報受容蛋白質フィトクロムの分子構造" 組織培養(ニュ-サイエンス社). 17. 425-430 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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