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滑り運動系の微細構造

Research Project

Project/Area Number 02239105
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionOkazaki National Research Institutes

Principal Investigator

月田 承一郎  岡崎国立共同研究機構, 生理学研究所, 教授 (50155347)

Co-Investigator(Kenkyū-buntansha) 若林 克三  大阪大学, 基礎工学部, 助手 (00029521)
盛田 フミ  北海道大学, 理学部, 教授 (80000818)
八木 直人  東北大学, 医学部, 助手 (80133940)
沼田 治  筑波大学, 生物科学系, 講師 (50189354)
岡本 洋  順天堂大学, 医学部, 講師 (70103877)
Project Period (FY) 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥28,000,000 (Direct Cost: ¥28,000,000)
Fiscal Year 1990: ¥28,000,000 (Direct Cost: ¥28,000,000)
Keywords滑り運動 / アクチン / ミオシン / 電子顕微鏡 / 急速凍結 / X線回折
Research Abstract

昨年度の成果を踏まえて,アクチンミオシン系の滑り運動系の動的微細構造の解析を進めた。本年度の本研究班の成果のうち,最も注目すべきものは,Caged化合物を用いた急速凍結電子顕微鏡法の完成と,その応用である。以下に本年度の成果をまとめる。
1.Caged化合物を用いた急速凍結電子顕微鏡法の開発(月田承一郎):昨年度より開発をはじめていたが,本年度はすべての技術的問題点を克服し,ほぼ完成した。この凍結システムを用いて2つの実験を行ないつつある。 第一は,Caged ATPを利用して、アクトS1複合体から,S1が解離していく様子を,ミリ秒時間分解能のコマ取り電子顕微鏡像として追跡していこうとするものである。多くの努力の結果,S1がアクチンフィラメントからその結合様式を変えながら離れていく様子をとらえることに成功した。その像の解釈は,これから慎重に行なわれなくてはならないが,滑り運動の分子機構を考える上で,極めて貴重なデ-タが得られたものと思われる。第二の実験は,やはりCaged ATPを利用して,本当に最大収縮速度で収縮(短縮)している途中のミオシンクロスブリッジの姿をとらえようとするものである。 現在,そのような姿がとらえはじめており,最終年度の解析が期待できる。
2.ミオシン分子上のアクチン結合部位が詳しく解析された(山本・盛田)
3.酵母を用いて,種々の遺伝子組換えアクチンを作製する道を初めて開いた(沼田治)。
4.収縮(短縮)中の骨格筋からX線回折パタ-ンを得ることに成功しつつある(八木直人・若林克三)。
5.月田らとは異なった方向から,Caged化合物と電顕像を結びつける試みを行ない,見通しがたつようになった(徳永万喜洋)。

Report

(1 results)
  • 1990 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [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] Tokunaga,M: "Structure and structural changes of the myosin head." Advances in Biophysics.

    • Related Report
      1990 Annual Research Report
  • [Publications] Yamamoto,K.: "Shift of binding site at the interface between actin and myosin." Biochemistry. 29. 844-848 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yamamoto,K.: "Identification of the site important for the actinーactivated MgATPase activity of myosin subfragmentー1." J.Mol.Biol.217. 229-233 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hirono,M.: "“Tetrahymena actin":Copolymerization with skeletal muscle actin and interactions with muscle actinーbinding proteins." J.Biochem.107. 32-36 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Edamatsu,M.: "Purification and characterization of Tetrahymena profilin." B.B.R.C.170. 957-962 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Matsubara,I.: "Crossーbridge movement in fast and slow skeletal muscles of the chick." J.Physiol.

    • Related Report
      1990 Annual Research Report
  • [Publications] Suzuki,R.: "Inhibition of actomyosin subfragment 1 ATPase activity by analog peptide of the actinーbinding site around the Cys(SH1) of myosin heavy chain" J.Biol.Chem.265. 4939-4943 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Inoue,K.: "Caーdependent protein phosphatase which dephosphorylates regulatory light chainーa in scallop smooth muscle myosin." J.Biochem.107. 872-878 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Eto,M.: "Roles of the amino acid side chains in the actinーbinding Sーsite of myosin heavy chain." J.Biochem.108. 499-504 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hasegawa,Y.: "Regulatory muchanism by the phosphorylation of 20ーkDa light chain of porcine aorta smooth muscle myosin." J.Biochem.108. 909-913 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Tregear,R.T.: "Xーray diffraction and electron microscopy from Lethocerus flight muscle partially relaxed by Adenylylimidodiphosphate and ethylene glycol." J.Mol.Biol.214. 129-141 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Konishi,M.: "Timeーresolved Xーray differaction studies of a single frog skeletal fiber." Biophys.Chem.

    • Related Report
      1990 Annual Research Report
  • [Publications] Wakabayashi,K.: "Progress in Xーray synchrotron diffraction studies of muscle contraction." Synchrotron Radiation Handbook.

    • Related Report
      1990 Annual Research Report

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

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