CORE--OPTICAL TRAPPING AND MICROSCOPY

这个核心的主要目标是为新的光学设备提供一种设施 向本计划项目的研究人员提供仪器。初级阶段 核心的功能将是协助需要 专业知识和专门设备在个别分部中不可用 项目实验室。核心成员将提供指导和 为项目不同成员在使用中提供专家帮助 光学捕获和全内反射偏振荧光 显微镜(TIRPFM)。这种最先进的共享设施提供了 GB/T1149.1-1989电机单机机械性能测试方法 蛋白质与与之相连的荧光探针的空间定位 单个工程蛋白的亚结构域。核心将进一步 开发这些技术以更好地控制蛋白质的位置 三维分子，并提高时间分辨率 测量单分子通过时的取向 它们的酶周期。单个正常和工程化细胞的体外分析 马达分子将毫不含糊地测量基本跨桥 特性，如刚度、方向变化、单向力、步长 长度和动力学作为应变的函数，并将这些特性与 这种蛋白质的结构特征。

The major aim of this Core is to provide a facility for novel optical instrumentation to the investigators of this Program Project. The primary function of the Core will be to assist in experiments that require expertise and specialized equipment not available in the individual sub- project laboratories. The members of the Core will provide guidance and expert help to the different members of the Program Project in using Optical Trapping and Total Internal Reflection Polarized Fluorescence Microscopy (TIRPFM). This state of the art shared facility provides methods for measuring the mechanical properties of individual motor proteins and the spatial orientation of fluorescent probes linked to subdomains of individual engineered proteins. The core will further develop these techniques to better control the position of protein molecules in three dimensions and to improve the time resolution for measuring the orientation of single molecules as they proceed through their enzymatic cycles. In vitro analysis of single normal and engineered motor molecules will unambiguously measure fundamental cross-bridge properties such as stiffness, orientation changes, unitary force, step length and kinetics as a function of strain and relate these properties to structural features of this protein.