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

我们的目标是在蛋白质原子结构的基础上理解马达蛋白的分子机制。我们描述了我们的结果，从这个赠款如下。(1)We鉴定了肌球蛋白N-末端在肌动球蛋白刚性复合物中的位置。(2)We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酶在调节Ca.（3）甲醇捕获处于关闭状态的细丝。（4）应用电子断层扫描技术重建肌横桥的三维结构。(5)We第一次通过冷冻电子显微镜在溶液中观察到单个肌球蛋白分子。(6)We在导通状态下获得高分辨率的细丝结构。(7)We重建松弛状态下含有粗丝的肌球蛋白的三维结构。(8)我们开发了一个新的环境来开发新的图像分析技术。（9）首次可视化了微管蛋白-驱动蛋白复合物的三维结构。（10）我们检测到了细纤维的结构变化，这是由Ca的加入引起的。（11）我们通过细丝的二维晶体观察肌钙蛋白。（12）我们用冷冻电镜鉴定了nebulin在天然细丝中的位置。（13）在强直复合体中检测到肌球蛋白的波动部分。（14）冷冻电镜观察肌动蛋白波动模式的变化。

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.