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X-ray Study of Actin Monomer-Myosin Complex and ESR Angular Measurement of Alkaline Light Chains

Research Project

Project/Area Number 10680633
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionOsaka University

Principal Investigator

ARATA Toshiaki  Graduate School of Science, Osaka Univ. Research Associate, 大学院・理学研究科, 助教授 (70151165)

Co-Investigator(Kenkyū-buntansha) KATSUZOU Wakabayashi  Graduate School of Engineering Science, Osaka Univ.Professor, 大学院・基礎工学研究科, 教授 (00029521)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2000: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordsmonomeric actin / myosin head / kinesin / tubulin / small-angle x-ray scattering / electron spin resonance / crystallography / spin label / 非重合アクチン / アクトミオシン
Research Abstract

We made a nonpolymerizable G-actin by MBS and DHT and measured X-ray solution scattering from a 1 : 1 acto-myosin S1 complex. The radius of gyration (Rg) of the complex was 49 A and apparent molecular weight was 165 kD.The model consisted of atomic structures of actin and S1 which fits the whole scattering curve closely was searched on the basis of rigid-body rotation and translation of two molecules. The uniqueness of the model was checked by a complex of an asymmetric DNaseI-actin and S1. A tip of S1 binds to actin subdomain 1 as expected from the previous electron microscopy of a myosin molecule whose heads are decorated with a monomeric G-actin (J.Struct.Biol. ('98)123, 8). In the presence of ADP, Rg decreased by 3 A with no change in molecular weight. The best fit model was obtained by twisting the two molecules each other through nearly 90 degrees about a long axis of S1 with retaining their main binding site. The complex of dimeric or timeric actin and S1 is now examined, and crystallization is in progress. The computer-fitting of ESR spectra from spin-labeled myosin light-chain in muscle fibres was done by a combination of two equimolar Gaussian distributions. The spectrum of relaxed muscle fibres was fitted with an asymmetric distribution and resolved into two broad angular distributions, suggesting that a pair of the myosin light-chain domains (neck regions) appear fixed loosely at different orientations on the myosin filament backbone. The spectrum from active muscle fibres is now analyzed. The tubulin-binding region of kinesin was spin-labeled using site-directed mutagenesis. A side-chain mobility estimated from spin-label ESR was restricted by tubulin binding, The immobilization depends on the occupancy of kinesin on a microtubule, suggesting a propagation of structural change along microtubule.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Kim,D.-S.: "X-ray Diffraction Studies on the Structural Changes of Rigor Muscles Induced by Binding of Phosphate Analogs in the Presence of MgADP"Biophys.Chem.,. 74. 71-82 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata,T.: "Electron Microscopic Observation of Monomeric Actin Attached to a Myosin Head"J.Struct.Biol.. 123. 8-16 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata,T.: "Structure of the Monomeric Actin-Myosin Head Complex as Revealed by X-ray Solution Scattering"Adv.Exp.Med.Biol.. 453. 73-78 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takezawa,Y.: "Backward Movements of Cross-Bridges by Application of Stretch and by Binding of MgADP to Skeletal Muscle Fibers in the Rigor State as Studied by X-ray Diffraction"Biophys.J.. 76. 1770-1783 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 荒田敏昭: "筋肉モータータンパク質アクチンとミオシンのダイナミックな分子構造と相互作用(ミニレビュー)"生化学. 72. 388-392 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata,T.: "Current Methods in Muscle Physiology. Chapter9.The use of spin probes"Oxford Univ.Press.. 223-239 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kim, D.-S., Takezawa, Y., Ogino, M., Kobayashi, T., Arata, T.and Wakabayashi, K.: "X-ray Diffraction Studies on the Structural Changes of Rigor Muscles Induced by Binding of Phosphate Analogs in the Presence of MgADP"Biophys.Chem.. 74. 71-82 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata, T.: "Electron Microscopic Observation of Monomeric Actin Attached to a Myosin Head."J.Struct.Biol. 123. 8-16 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata, T., Kimura, S., Sugimoto, Y., Takezawa, Y., Iwasaki, N.and Wakabayashi, K.: "Structure of the Monomeric Actin-Myosin Head Complex as Revealed by X-ray Solution Scattering"Adv.Exp.Med.Biol. 453. 73-78 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takezawa, Y., Kim, D.-S., Ogino, M., Sugimoto, Y., Kobayashi, T., Arata, T.& Wakabayashi, K: "Backward Movements of Cross-Bridges by Application of Stretch and by Binding of MgADP to Skeletal Muscle Fibers in the Rigor State as Studied by X-ray Diffraction"Biophys.J.. 76. 1770-1783 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata, T: "Dynamic Structure and Interaction of Actin-Myosin complex (in Japanese)"Seikagaku. 72(5). 388-392 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Arata, T: "The use of spin probes. In Current Methods in Muscle Physiology"Oxford Univ.Press. 223-239 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 荒田敏昭: "筋肉モータータンパク質アクチンとミオシンのダイナミックな分子構造と相互作用"生化学. 72巻5号. 388-392 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Arata: "Angular Distribution of Spin-Labeled Light Chain of Myosin Heads in Muscle Figres, as Derived from Close Fit of ESR Spectrum"Journal of Muscle Res.Cell Motil.. 21(印刷中). (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Takezawa: "Backward Movement of Cross-bridges by Application of Stretch and by Binding of MgADP to Skeletal Muscle Fibers in the Rigor State as Studied by X-ray Diffraction"Biophysical Journal. 76. 1770-1783 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 荒田敏昭: "筋肉モーター蛋白質アクチンとミオシンのダイナミックな分子構造と相互作用"生化学. 72巻5号(印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] T.Arata: "X-ray Solution Scattering study of the actin dimer-myosin head complex"J. Muscle Res. Cell Motil.. 20. 325-327 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] T.Arata: "Electron Microscopic Observation of Monomeric Actin Attached to a Myosin Head" Journal of Structural Biology. 123. 8-16 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] D.-S.kim: "X-ray Dittraction Studies on the Structural Changes of Rigor Muscles Induced by Binding of Phosphate Analogs in the Presence of MgADP" Biophysical Chemistry. 74. 71-82 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Arata: "Stucure of the Monomeric Action-myosin Head Complex as Revealed by X-ray Solution Scattering" Adv.Exp.Med.Biol.453. 73-78 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Takezawa: "Backward Movements of Cross-Bridges by Application of Stretch and by Binding of MgADP to Skeletal Muscle Fibers in the Rigor State as Studied by X-ray Diffraction" Biophysical Journal. 76(印刷中). (1999)

    • Related Report
      1998 Annual Research Report

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

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