STRUCTURE, FUNCTION & MECHANISM OF CONTRACTILE PROTEINS

该计划项目（宾夕法尼亚肌肉研究所）的职能， 分子生物学跨学科合作研究计划 肌肉收缩和肌球蛋白为基础的细胞运动机制。主要 该计划的主题是结构生物学，机械化学， 收缩蛋白的蛋白质设计和序列特异性。它 整合了从基因表达，异构体多样性， 酶活性、分子运动和反应动力学 收缩蛋白分子完全构成肌原纤维。客观 是要了解这些蛋白质，单独和组织的， 特定的大分子集合体，执行它们的化学任务， 分子水平上的结构和机械能转换。的 项目内的项目和调查人员是相互依存的， 通过互补的研究目标紧密联系在一起， 技术、资源和培训计划。新型仪器将成为 为研究开发的将包括激光光学陷阱，测定 力、位移距离和两个单畴的畴间运动 分子和收缩蛋白质的有序阵列，时间分辨 荧光偏振显微镜和超快速冷冻电子显微镜 与瞬时分子事件同步的显微镜。这些瞬变 将通过“笼状”基质的激光脉冲光解引发， 信号分子和快速的机械扰动。小说 技术将包括新的真核共表达系统， 结构-功能研究，截短的、突变的或 嵌合蛋白质在遗传上可接受的生物体中，双功能 用于定量蛋白质取向动力学的荧光标记， 单个肌原纤维的力学分析。来自特殊组织的肌球蛋白， 如平滑肌和振荡肌，将得到深入研究。的 长期科学目标和计划开发的技术 项目旨在了解正常的组装和功能， 以及收缩蛋白的病理生理学相关的， 心血管和肺系统。

The Program Project (Pennsylvania Muscle Institute) functions as in interdisciplinary program for collaborative studies on the molecular mechanism of muscle contraction and myosin-based cell motility. The major themes of the Program are the structural biology, mechano-chemistry, protein design and sequence specificity of the contractile proteins. It integrates investigations from genetic expression, isoform diversity, enzymatic activity, molecular motions and reaction kinetics in isolated contractile protein molecules fully constituted myofibrils. The objective is to understand how these proteins, individually and organized in highly specific, macromolecular assemblies, perform their task of chemical, structural and mechanical energy transduction at the molecular level. The projects and investigators within the Program are interdependent and closely linked through complementary research goals and shared technologies, resources and training programs. Novel instrumentation to be developed for the research will include laser optical traps, determination of force, translocation distance and inter-domain motions of both single molecules and ordered arrays of contractile proteins, time-resolved fluorescence polarization microscopy, and ultra-rapid freezing electron microscopy synchronized to transient molecular events. These transients will be initiated by laser pulse photolysis of "caged" substrate or signaling molecules and by rapid mechanical perturbations. Novel techniques will include new eukaryotic co-expression systems for protein structure-function studies, transgenic expression of truncated, mutated or chimeric proteins in genetically amenable organisms, bifunctional fluorescence labeling for quantitative protein orientational dynamics and mechanical analysis of single myofibrils. Myosin from specialized tissues, such as smooth and oscillatory muscles, will be studied intensively. The long-term scientific aims and technologies developed in the Program Project are directed to understanding the normal assembly and function as well as pathophysiology of contractile proteins relevant to the cardiovascular and pulmonary systems.