权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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。我们在下面描述我们的结果。(1)我们确定了肌凝蛋白n端在肌动肌凝蛋白复合体中的位置。(2)我们确定了肌动蛋白上的原肌球蛋白结合位点，这影响了肌动蛋白激活的atp酶在钙调节中的协同性。(3)甲醇使细丝处于关闭状态。(4)通过电子断层扫描重建收缩肌的三维交叉桥结构。(5)首次用低温电镜观察了溶液中单个肌球蛋白分子。(6)在导通态上获得了细细丝的高分辨率结构。(7)在松弛状态下重建了含有粗丝的肌球蛋白的三维结构。我们开发了一个新的环境来开发新的图像分析技术。(9)我们首次可视化了微管蛋白-激酶复合物的三维结构。（10）我们检测到了Ca的加入引起的细丝的结构变化。（11）我们通过细丝的二维晶体观察到了肌钙蛋白。（12）利用电子冷冻显微镜鉴定了星云在天然细丝中的位置。（13）我们在刚性复合体中检测到肌球蛋白的波动部分。（14）通过电子冷冻显微镜检测肌动蛋白波动模式的变化。