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A Rapid-freeze Electron Microscopy in Biological Materials

Research Project

Project/Area Number 01480541
Research Category

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

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionOkazaki National Research Institutes (1990)
Tokyo Metropolitan Organization for Medical Research (1989)

Principal Investigator

TSUKITA Shoichiro  Okasaki National Research Institutes, Prof., 生理学研究所, 教授 (50155347)

Co-Investigator(Kenkyū-buntansha) TSUKITA Sachiko  Okazaki National Research Institute, assistant Prof., 生理学研究所, 助手 (00188517)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1989: ¥4,500,000 (Direct Cost: ¥4,500,000)
KeywordsCaged compounds / Electron Microscope / Myosin / Rapid freezing / Liquid helium / ATP / 急速凍結法 / 骨格筋 / カルシウム / 蛋白質
Research Abstract

The interaction between myosin subfragment 1(Sl)and actin filaments after the photolysis of P^3-1-(2-nitrophenyl)ethyl ester of ATP(caged ATP)was analyzed using a newly-developed freezing system with liquid helium. Actin and Sl(100g mu each)formed a rope-like double helix characteristic of rigor in the presence of 5 mM caged ATP at room temperature. A UV flash with a half duration time of 0.3 ms photoreleased -0.8 mM of ATP. At 15 ms after photolysis, the rope-like double helix was partially disintegrated. S1 on actin filaments gradually decreased in number up to 35 ms after photolysis and their association was in a somewhat cooperative manner. No more changes of Sl attached on actin filaments were detected from 35 to 200 ms. At 1 min after photolysis, ATP was consumed and the rope-like double helix was reformed. Taking recent analyses of actomyosin kinetics into consideration, we concluded that most Sl observed on actinfilaments at 15-200 ms are so called "weakly-bound Sl"(SL. ATP or Sl. ADP. Pi)and that the gradual decrease of the weakly-bound Sl during 15-35 ms is due to the transition from the rapid equilibrium of actin. Sl. ATP/SI. ATP to that of actin. Sl. ADP. Pi/Sl. ADP. Pi.

Report

(3 results)
  • 1991 Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Tsukita,Sh.,Tsukita,Sa.: "Isolation of cell-to-cell adherens junction from rat liver." J.Cell Biol.108. 31-41 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita,Sa.,Hieda,Y.,Tsukita,Sh.: "A new 82-kD barbed end-capping protein(redixin)localized in the cell-to-cell adherens junction:Purification and characterization" J.Cell Biol.108. 2369-2382 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hieda,Y.,Tsukita,Sa.,Tsukita,Sh.: "A new high molecular mass protein showing unique localization in desmosomal plaque." J.Cell Biol.109. 1511-1518 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita,Sh.,Itoh,M.,Tsukita,Sa.: "A new 400 kD-protein from isolated adherens junctions:Its localization at the undercoat of adherens junctions and at microfilament bundles such as stress fibers and circumferential bundles." J.Cell Biol.109. 2905-2915 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Matsumoto,G.,Tsukita,Sh.,Arai,T.: "Organization of the axonal cytoskeleton:Differentiation of the microtubule and actin filament arrays." In Cell Movement.Alan R.Liss Inc.335-356 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita,Sh.: "The undercoat of adherens junctions:A Key specialized structure in organogenesis and carcinogenesis." Cell Struct.Funct.15. 7-12 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita, Sh. and Tsukita, Sa: "Isolation of cell-to-cell adherens junction from rat liver." J. Cell Biol.108. 31-41 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita, Sa., Hieda, Y. and Tsukita, Sh.: "A new 82-kD barbed end-capping protein (radixin) localized in the cell-to-cell adherens junction : Purification and characterization." J. Cell Biol.108. 2369-2382 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hieda, Y., Tsukita, Sa. and Tsukita, Sh.: "A new high molecular mass protein showing unique localization in desmosomal plaque." J. Cell Biol.109. 1511-1518 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukika. Sh. et al.: "A new 400kD-protein from isolated adherens junctions : Its localization at the undercoat of adherens junctions and at microfilament bundles such as stress fibers and circumferential bundles." J. Cell Biol.109. 2905-2915 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Higuchi, M., Mitsuno, T., Sugimoto, M., Okamaoto, A., Hirose, S., Tsukuta, Sh. and Osawa, T.: "Tumoricidal activitiy of lymphotoxin (tumor necrosis factor) in vivo : Its effects on macrophages." J. Biological Response Modifiers. 7. 619-630 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita, Sh., Tsukita, Sa. and Nagafuchi, A.: "The undercoat of adherens junctions : A key specialized structure in organogenesis and carcinogensis." Cell Struct. Funct.15. 7-12 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nishikawa, S., Tsukita, Sa., Tsukita, Sh. and Sasa, S.: "Localization of adherens junction proteins along the possible sliding interface between secretory ameloblasts of the rat incisor." Cell Struct. Funct.15. 245-250 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tsukita,Sh.: "The undercoat of adherens junctions:A key specialized structure in organogenesis and carcinogenesis." Cell Struct.Funct.15. 7-12 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Nishikawa,S.: "Localization of adherens junction proteins along the possible sliding interface between secretory ameloblasts of the rat incisor." Cell Struct.Funct.15. 245-250 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Sato,N.: "Radixin,a barbedーendーcapping actinーmodulating protein,is concentrated at the cleavage furrow during cytokinesis." J.Cell Biol.

    • Related Report
      1990 Annual Research Report
  • [Publications] Muto,E.: "Doubleーrowed organization of inner dynein arms in Chlamydomonas flagella revealed by tiltーseries thinーsection electron microscopy." J.Cell Sci.

    • Related Report
      1990 Annual Research Report
  • [Publications] Tsukita,Sa: "Specific protoーoncogenic tyrosine kinases of src family are enriched in cellーtoーcell adherens junctions where the level of tyrosine phosphorylation is elevated." J.Cell Biol.

    • Related Report
      1990 Annual Research Report
  • [Publications] 月田承一郎,月田早智子: "Isolation of cell-to-cell adherens junction from rat liver." J.Cell Biol.108. 31-41 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 月田早智子,檜枝洋記,月田承一郎: "A new 82-kD barbed end-capping protein(redixin)localized in the cell-to-cell adherens junction:purification and characterization" J.Cell Biol.108. 2369-2382 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 檜枝洋記,月田早智子,月田承一郎: "A new high molecular mass protein showing unique localization in desmosomal plaque." J.Cell Biol.109. 1511-1518 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 月田承一郎,伊藤雅彦,月田早智子: "A new 400 kD-protein from isolated adherens junctions:Its localization at the undercoat of adherens junctions and at microfilament bundles such as stress fibers and circumferential bundles." J.Cell Biol.109. 2905-2915 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 松本元,月田承一郎,新井孝夫: "Organization of the axonal cytoskeleton:Differentiation of the microtubule and actin filament arrays." In Cell Movement.Alan R.Liss Inc.335-356 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 月田承一郎,月田早智子,永渕昭良: "The undercoat of adherens junctions:A key specialized structure in organogenesis and carcinogenesis." Cell Struct.Funct.

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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