Development of observation technology with electron microscope of force-generating myosin

我们试图开发产生肌球蛋白的电子显微镜观察技术。一项必要的技术是观察肌球蛋白细丝和肌动蛋白细丝在力产生过程中的主动相互作用。成功的话，肌动蛋白细丝可以被磁珠拉动，磁珠与肌动蛋白共价连接。另一种是用电子冷冻显微镜观察单个肌球蛋白头部和肌动蛋白细丝的三维结构。我们开发了用于冷冻水化标本的单颗粒分析和电子断层扫描的新的三维重建程序，并获得了高分辨率的肌动蛋白细丝/肌球蛋白复合体的3D图和单头肌球蛋白与单个肌动蛋白细丝结合的初步3D图。

We tried to develop observation technology with electron microscopes of force generating myosin. One necessary technology is to observe the active interaction between a myosin filament and an actin filament during force generation. The actin filament could be, in success, pulled by a magnetic bead, which was linked to the actin covalently. The other is to observe three-dimensional structure of single myosin head and actin filament by electron cryo-microscopy. We developed new three-dimensional reconstruction programs for single particle analysis and electron tomography of frozen-hydrated specimens and obtained highly-resolved 3D map of actin filaments/actomyosin complexes and preliminary 3D maps of single head myosin bound to a single actin filament.