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Identification of protein sequence which bind to phytochrome A.

Research Project

Project/Area Number 07839015
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section時限
Research Field 光生物学
Research InstitutionYokohama City University

Principal Investigator

MANABE Katsushi  Yokohama City University, Faculty of Science, Professor, 理学部, 教授 (10112484)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
Keywordsphytochrome / chromophore-protein interaction / phage library / epitope / リセプタータンパク質 / アフィニティー分画 / ファージディスプレイ / 抗体 / 化学架橋
Research Abstract

A monoclonal antibody designated Mep-1, was raised against purified phytochrome A from pea. The binding of this antibody (class IgG1) to partially degraded phytochrome (114 kDa) caused a considerable increase in the far-red peak at the stationary state after red-light-irradiation. The effect reached a plateau value when the antibody and phytochrome were present in approximately equimolar amounts. The dark transformation of Pfr to Pr of the 114-kDa phytochrome was inhibited by the addition of the antibody. However, binding of the antibody to the undegraded 121-kDa phytochrome had no effects on the spectrum of the red-light-induced photostationary state. The site at which the antibody bound to phytochrome was determined to be between amino acid residues 256 and 383 of pea phytochrome A by peptide mapping. The site of the binding was further studied using phage random peptide display system and was determined to be residues from D-354 to L-357 (DAVL). A synthetic octa-peptide SADAVLPQ (corresponding to residues from 352 to 359 of the phytochrome) indeed had strong binding activity to Mep-1. The importance of the site for the chromophore-apoprotein interaction is discussed.
Experiments to find peptide sequence which specifically bind to the physiologically active Pfr form of phytochrome are in progress using the phage random peptide display library.

Report

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

    (12 results)

All Other

All Publications (12 results)

  • [Publications] 眞鍋勝司,中澤美紀: "植物の光形態形成の光受容色素フィトクロムの機能部域。" 横浜市立大学論叢自然科学系列. 6(1). 39-51 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Iwakami.S.et al.: "Spectral properties and photoactivities of intermediates of photoconversion from the red-light-absorbing to far-red-light-absorbing form of pea phytochrome." J.Photochem.Photobiol.B.33. 239-244 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nakazawa,M.et al.: "A monoclonal antibody affects chromophore apoprotein interactions of phytochrome A." Photochem.Photobiol.65. in press (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Manabe,K.,Nakazawa,M.: "The structure and function of phytochrome A:the roles of entire molecule and of its various parts." J.Plant Res.110. in press (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 中澤美紀,眞鍋勝司: "現代化学増刊30「植物のシグナルトランスダクション(蓮沼,平野編)東京化学同人" 遺伝子導入による機能領域の解析。, 6 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Iwakami, S., Yoshizawa, N., Hamaguchi, H., Inoue, Y., Manabe, K.: "Spectral properties and photoactivities of intermediates of photoconversion from the red-light-absorbing to far-red-light-absorbing form of pea phytochrome" J.Photochem.Photobiol.B.33. 239-244 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nakazawa, M., Moriuchi, N., Wada, N., Hirose, T., Tokutomi, S., Manabe, K.: "A monoclonal antibody affects chromophore apoprotein interactions of phytochrome A" Photochem.Photobiol.65 (in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Manabe, K., Nakazawa, M.: "The structure and function of phytochrome A : the roles of entire molecule and of its various parts" J.Plant Res.110 (in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nakazawa,M.et al. :"A monoclonal antibody affects chromophore apoprotein interactions of phytochrome A." Photochem.Photobiol.65(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Manabe,K.,Nakazawa,M. :"The structure and tunction of phytochrome A : the roles of entire molecule and of its various parts." J.Plant Res.110(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 眞鍋勝司,中澤美紀;: "植物の光形態形成の光受容色素フィトクロムの機能部域。" 横浜市立大学論叢自然科学系列. 6. 39-51 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 中澤美紀,眞鍋勝司:"現代化学増刊30「植物のシグナルトランスダクション (蓮沼,平野編)東京化学同人" 遺伝子導入による機能領域の解析。, 6 (1996)

    • Related Report
      1996 Annual Research Report

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

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