Mechanism of protein motor based on molecular structure using protein-engineering and cryo-electron microscopy

利用蛋白质工程和冷冻电子显微镜研究基于分子结构的蛋白质马达机制

基本信息

批准号：
06454663
负责人：
WAKABAYASHI Takeyuki
金额：
$ 4.22万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

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.

我们的目标是在蛋白质原子结构的基础上了解运动蛋白的分子机制。我们在下面描述了这项资助的结果。(1)我们确定了肌球蛋白 N 末端在肌动球蛋白严格复合物中的位置。 (2)我们鉴定了肌动蛋白上的原肌球蛋白结合位点，该位点影响肌动蛋白激活的 ATP 酶在 Ca 调节中的协同作用。 (3)甲醇在关闭状态下捕获细丝。 (4)利用电子断层扫描技术重建收缩肌横桥的三维结构。 (5)我们首次通过冷冻电子显微镜观察到溶液中的单个肌球蛋白分子。 (6)我们在导通状态下获得了细丝的高分辨率结构。 (7)我们在松弛状态下重建了含有粗丝的肌球蛋白的三维结构。(8)我们开发了新的环境来开发新的图像分析技术。 (9)我们首次将微管蛋白-驱动蛋白复合物的三维结构可视化。 (10)我们检测到了细丝的结构变化，这是由Ca的添加引起的。(11)我们通过细丝的二维晶体观察了肌钙蛋白。 (12)我们通过电子冷冻显微镜确定了星云蛋白在天然细丝中的位置。 (13)我们在僵直复合体中检测到肌球蛋白的波动部分。 (14)我们通过电子冷冻显微镜检测了肌动蛋白波动模式的变化。