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THE STRUCTURAL ANALYSIS OF PHOTO-TRANSDUCTION PROTEINS

Research Project

Project/Area Number 10044264
Research Category

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

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionNAGOYA UNIVERSITY

Principal Investigator

USUKURA Jiro  NAGOYA UNIVERSITY,SCHOOL OF MEDICINE., ASSOCIATE PROFESSOR, 医学部, 教授 (30143415)

Co-Investigator(Kenkyū-buntansha) NISHIZAWA Yuji  SCHOOL OF MEDICINE., RESEARCH ASSOCIATE, 医学部, 助手 (80252229)
WAKABAYASHI Takashi  SCHOOL OF MEDICINE., PROFESSOR, 医学部, 教授 (00079998)
松本 博行  Oklahoma State University, Medical School, 准教授
篠原 利道  Harvard Unicersity, Medical School, 准教授
山崎 哲雄  Wayne State University, Kresge Eye Instit, 教授
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsPhototransduction / Photoreceptor / EELS / Phosphodiesterase / Electron microscopy / cGMP / synapse / Ca / トランスミッター / カベオリン / cGMP分解酵素 / 蛋白質複合体 / 構造解析 / 低角度回転蒸着法
Research Abstract

The aim of this international cooperative research with Dr Akio Yamazaki, Kresge Eye Institute, Wayne State University, was to reveal morphological counterpart of the photo-transduction process in the vertebrate photoreceptor cells. In 1998, we have investigated morphological change of phosphodiesterase (PDE), one of the key components in phototransduction, accompanied by functional changes in PDE subunits and to elucidate new roles of PDE in visual signal transduction mechanism. In contrast to significant progress in understanding the flow of signals in phototransduction and the roles of each component in the flow of signals, the relationship in PDE subunits is unknown because of difficulty of purification and expression of intact Pαβ and lack of information about regulation of Pαβ-Pγinteractions. However, in this study, Pαβ-Pγ interactions were partially elucidated with new findings on the mechanism of Pγ modifications and unexpected finding on the PDE role in the increase of cytoplasmic cGMP level in rod outer segments. Our recent development in the isolation and purification of bovine Pαβ with or without Pγ also made possible to observe the molecules under electron microscope. All types of PDE (αβγγ, αβγ, αβ) consisted basically of two crooked strands with characteristic shape that were arranged as making a ring. Suggestible structural changes induced by dislocation of Pγ from complete PDE were arrested as three dimensional shift of molecules in position. In 1999, we started to study the functional structure of synaptic ribbons that are located at the active site and surrounded by a halo of synaptic vesicles. Our new electron microscopy revealed surprisingly that synaptic ribbon was associated with Ca. Since synaptic ribbon represented abundant phosphorus element signals in electron energy loss spectral imaging, it is clear that the synaptic ribbon is a derivative of the membrane.

Report

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

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Seno K.: "A possible role of RGS 9 in phototransduction : A bridge between the cGMP-phosphodiesterase system and the guanylyl cyclase system"J. Biol. Chem.. 273. 22169-22172 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kachi, S.: "Detailed localization of photoreceptor guanylate cyclase activating protein-1 and -2 in mammalian retainas using light and electron microscopy"Exp. Eye Res.. 68. 465-473 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Teranishi Ma.: "Effects of coenzyme Q10 on changes in the membrane potential and rate of generation of reactive oxygen species in hydrazine- and chloramphenicol-treated rat iver mitochondria"Arch. Biochem. Biophys.. 366. 157-167 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sato A.: "Inhibitor peptide SNP-1 binds to a soluble form of BST-1/CD157 at a 2:2 stoichiometry"Eur. J. Biochem.. 264. 439-445 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nishizawa Y.: "Initiating ocular proteomics for cataloging bovineretinal proteins : microanalytical techniques permit the identification of proteins derived from a novel photoreceptorpreparation"Exp. Eye Res.. 69. 195-212 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe N.: "Perciliary structure of developing rat photoreceptor cells. A deep etch replica and freeze substitution study"J. Electron Microsc.. 48. 929-935 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Seno K., Kishigami A., Ihara S., Maeda T., Bondarenko V.A., Nishizawa, Y., Usukura J., Yamazaki A. & Hayashi F.: "A possible role of RGS 9 in phototransduction : A bridge between the cGMP-phosphodiesterase system and the guanylyl cyclase system"J. Biol. Chem. 273. 22169-22172 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kachi S. Nishizawa Y. Olshevskaya E. Yamazaki A. Miyake Y. Wakabayashi T. Dizhoor A. & Usukura J.: "Detailed localization of photoreceptor guanylate cyclase activating protein-1 and -2 in mammalian retinas using light and electron microscopy"Exp. Eye Res.. 68. 465-473 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Teranishi Ma. Karbowski M. Kurono C. Nishizawa Y. Usukura J. Soji T. & Wakabayashi T.: "Effects of coenzyme Q10 on changes in the membrane potential and rate of generation of reactive oxygen species in hydrazine- and chloramphenicol-treated rat iver mitochondria"Arch. Biochem. Biopohys. 366. 157-167 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sato A. Yamamoto S. kajimura N. Oda M. Usukura J. Jingami H.: "Inhibitor peptide SNP-1 binds to a soluble form of BST-1/CD157 at a 2 : 2 stoichiometry"Eur. J. Biochem.. 264. 439-455 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nishizawa Y. Komori N. Usukura J. Jackson KW. Tobin SL. Matsumoto H.: "Initiating ocular proteomics for cataloging bovineretinal proteins : microanalytical techniques permit identification of proteins derived from a novel photoreceptor preparation"Exp. Eye Res.. 69. 195-212 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe N. Miyake Y. Wakabayashi T. Usukura J.: "Periciliary structure of development rat photoreceptor cells. A deep etch replica and freeze substitution study"J. Electron Microsc.. 48. 929-935 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nishizawa Y.: "Initiating ocular proteomics for cataloging bovine retinal proteins: microanalytical techniques permit the identification of proteins derived from a novel photoreceptor preparation."Exp.Eye Res.. 69. 195-212 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sato A.: "Inhibitor peptide SNP-1 binds to a soluble form of BST-1/CD157 at a 2:2 stoichiometry."Eur.J.Biochem.. 264. 439-445 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Watanabe N.: "Periciliary structure of developing rat photoreceptor cells.A deep etch replica and freeze substitution study."J.Electron Microsc.. 48. 929-935 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kachi S.: "Detalled localization of photoreceptor guanylate cyclase activating protein-1 and -2 in mammalian retinas using light and electron microscopy." Exp.Eye Res.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Seno K.: "A possible role of RGS 9 in phototransduction: A bridge between the cGMP-phosphodiesterase system and the guanvlvl cvclase system." J.Biol.Chem.273. 22169-22172 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hirako,Y.: "Cleavage of BP 180,a 180kDa bullous pemphigoid antigens,yields a 120 kDa collagenous extracellular polypeptide." J.Biol.Chem.273. 9711-971* (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Matsuhashi,T.: "Complete suppression of ethanol-induced formation of megamitochondria by 4-hydroxy-2,2,6,6-tetra-methyl-piperidine-1-oxyl(4-oh-TEMPO)." Free Radic.Biol.Med.24. 139-147 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 臼倉治郎: "バイオイメージング" 共立出版(株), 266 (1998)

    • Related Report
      1998 Annual Research Report

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

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