CORE--OPTICAL TRAPPING AND MICROSCOPY

该核心的主要目的是提供一个新的光学设施， 仪器仪表的研究人员这个计划项目。主 核心的功能将是协助进行需要 专业知识和专门设备，不具备在个别小组， 项目实验室。核心成员将提供指导， 专家帮助计划项目的不同成员使用 光阱和全内反射偏振荧光 显微镜检查（TIRPFM）。这种最先进的共享设施提供了 单台电动机机械性能测试方法 蛋白质和荧光探针的空间取向连接到 单个工程化蛋白质的亚结构域。核心将进一步 发展这些技术来更好地控制蛋白质的位置， 三维分子并提高时间分辨率 测量单个分子的方向 它们的酶循环体外分析单个正常和工程化的 马达分子将毫不含糊地测量基本的交叉桥 诸如刚度、取向变化、单一力、阶跃 长度和动力学作为应变的函数，并将这些性质与 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.