| 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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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | Muscle / Myosin / Actin / Tubulin / Kinesin / Electron microscopy / X-ray scattering / Site-directed mutagenesis / クライオ電子顕微鏡 / 画像解析 / カルシウム / 筋収縮 / 蛋白質工学 |
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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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