1995 Fiscal Year Final Research Report Summary
Mechanism of protein motor based on molecular structure using protein-engineering and cryo-electron microscopy
| Project/Area Number |
06454663
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biophysics
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| Research Institution | University of Tokyo |
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Principal Investigator |
WAKABAYASHI Takeyuki Univ.of Tokyo, School of Science, Projessor, 大学院・理学系研究科, 教授 (90011717)
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| Co-Investigator(Kenkyū-buntansha) |
YASUNAGA Takuo Univ.of Tokyo, School of Science, Research associate, 大学院・理学系研究科, 助手 (60251394)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Muscle / Myosin / Actin / Tubulin / Kinesin / Electron microscopy / X-ray scattering / Site-directed mutagenesis |
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| Research Abstract |
Our goal is to understand the molecular mechanism of motor proteins on the basis of atomic structure of proteins. We describe our result from this grant below.
(1)We identified the position of myosin N-terminus in actomyosin rigor complexes. (2)We identified the tropomyosin binding site(s) on actin, which affects cooperativity of actin-activated ATPase in regulation of Ca. (3)Methanol traps thin filaments on an off-state. (4)The three-dimensional structure of cross-bridge in contracting muscle was reconstructed by electron tomography. (5)We visualized single myosin molecules in solution by cryo-electron microscopy for the first time. (6)We obtained high resolutional structure of thin filaments on an on-state. (7)We reconstructed three-dimensional structure of myosin containing thick filaments on a relaxd state.(8) We developed a new environment to develop new image analysis techniques. (9)For the first time, we visualized three-dimensional structure of tubulin-kinesin complexes. (10)We detected the structural change of thin filaments, which is induced by the addition of Ca.(11)We observed troponin by two-dimensional crystal of thin filaments. (12)We identified the position of nebulin in native thin filaments by electron cryo-microscopy. (13)We detected fluctuating part of myosin in rigor complexes. (14)We detected the change of actin fluctuation mode by electron cryo-microscopy.
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[Publications] Kikkawa, M., Ishikawa, T., Miyazawa, A., Fujiyoshi, Y., Wakabayashi, T.& Hirokawa, N.: "Direct visualization of the microtubule lattice seam both in vitro and in vivo." J Cell Biol.127. 1965-1971 (1994)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kikkawa, M., Ishikawa, T., Miyazawa, A., Fujiyoshi, Y., Wakabayashi, T.& Hirokawa, N.: "Three-dimensional structure of tubulin-kinesin complex" Nature. 376. 274-277 (1995)
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「研究成果報告書概要(欧文)」より
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[Publications] Wakabayashi, T., Yasunaga, T., Miki, M., Saeki, K., Sutoh, & Sasaki, H.: "Structure and function of actin filaments revealed by sited-directed mutagenesis of actin and image , , analysis of cryo-electron micrographs" Structure and function of macromolecular assmbly. 244-252 (1997)
Description
「研究成果報告書概要(欧文)」より
