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Reconstitution of phototransduction channel from frog rod outer segments in planar lipid bilayers

Research Project

Project/Area Number 62570041
Research Category

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

Allocation TypeSingle-year Grants
Research Field General physiology
Research InstitutionNagoya University School of Medicine

Principal Investigator

SOKABE Masahiro  Nagoya University School of Medicine, Associate Prof., 医学部, 助教授 (10093428)

Co-Investigator(Kenkyū-buntansha) SHINOZAWA Takao  Faculty of Engineering,Gunma University,Associate Prof., 工学部, 助教授 (30025449)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1988: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1987: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsFrog / Photoreceptor / Cyclic GMP / Ion channel / Photoaffinity labeling / planar bilayer / Purification / 精製 / 再構成 / 抗体 / 光情報変換チャネル
Research Abstract

Vertebrate photoreceptors show hyperporalizing responses to light stimulations due to the closure of cGMP-dependent cation channels on outer segment membranes. The aim of this project is to identifiy and purify the cGMP-dependent ion channel protein. Integral membrane protein fraction from frog rod outer segments was labeled with [^3H]cGMP and four cGMP binding proteins with molecular weights of 250K, 100K, 92K, and 66K were identified. The incoropration of the fraction into planar lipid bilayers resulted in the appearance of at least three kinds of channel activity; two of them were related to cGMP. Since the 100K and 92K were phosphodiesterase, the 250K and/ar 66K proteins were strongly suggested to be cGMP dependent channel proteins. We performed partial purification of the 250K protein and succeeded in reconstituting of it into liposomes. By using polyclonal antibody (IGG) against the 250K protein the distribution of the protein was investagted by means of immuno-electronmicroscopy. The 250K protein seemed to be enriched on disc membranes rather than plasma membranes, which may be in harmony with recent reports indicating an enrichment of cGMP-dependent channels in disc membranes. Interestingly the IgG fraction reacts with both 250K and 66K proteins suggesting that the former is a precurser of the latter. It may also be possible these proteins are different types of cGMP-dependent channels. These problems would be solved by the further purification and reconstitution of these proteins.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] M.Tanifuji: J.Membr.Biol.99. 103-112 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Shinozawa: J.Biochem.102. 281-290 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 曽我部正博: 生物物理. 27. 21-30 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Sokabe: Brain Research. 443. 254-260 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y.Oosawa: Cell Structure & Function. 13. 51-60 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 曽我部正博: 蛋白質,核酸,酵素.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Yosizawa: "Retinal Proteins" VNU Science Press, 75-84 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 曽我部正博: "神経情報伝達分子" 培風館, 106-120 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Tanifuji: "An anion channel of sarcoplasmic reticulum incorporsted into planar lipid bilayers" J. Membr. Biol.99. 103-112 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T. Shinozawa: "Detection of cyclicGMP binding protein and the ion channel activity in frog rod outer segments" J. Biochem.102. 281-290 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Sokabe: "Reconstitution of ion channel into planar lipid bilayers" Biophysics. 27. 21-30 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Sokabe: "Effects of cyclic nucleotides and calcium on transduction and encoding processes in frog muscle spindle" Brain Research. 443. 254-260 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Sokabe: "Purification and recostitution of cyclicGMP dependent cation channel" Protein Nucleic Acid & Enzyme. in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T. Yoshizawa: Retinal Proteins. VNU Science Press, 75-84 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] N.Fujitsuka: Brain Research. 443. 243-253 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Sokabe: Brain Research. 443. 254-260 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Y.Oosawa: Cell Structure & Function. 13. 51-60 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Yanagisawa: Prog.Brain Res.74. 313-317 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Sokabe: Proc.2nd Int.Congr.CPB. 2. 815 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] 曽我部正博: 蛋白質、核酸、酵素.

    • Related Report
      1988 Annual Research Report
  • [Publications] F.Ito: "Mechanoreceptors" Plenum Press, 195-199 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] 曽我部正博: "神経情報伝達分子" 培風館, 106-120 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] T. Shinozawa: J. Biochemistry. 102. 281-290 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] T. Yoshizawa: Retinal Proteins(VNU Science Press). 75-84 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 曽我部正博: 生物物理. 27. 21-30 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Fujitsuka, N.: Brain Research. 443. 243-253 (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Sokabe, M.: Brain Research. 443. 254-260 (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yoshimura, A: Histochemistry.

    • Related Report
      1987 Annual Research Report
  • [Publications] 伊藤文雄: "生理学" 東西医学社, 566 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] 葛西道生: "神経系情報伝達分子" 培風館, 175 (1988)

    • Related Report
      1987 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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