Protein Structure and Dynamics from EPR Spectroscopy and MD Simulations

EPR 光谱和 MD 模拟的蛋白质结构和动力学

基本信息

批准号：
8277917
负责人：
TERRY P LYBRAND
金额：
$ 112.76万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2014-04-30
项目状态：
已结题

项目摘要

The long-term objectives of this program project are to develop computational tools that can be used to extract accurate structural and dynamical information from site-directed spin-labeling (SDSL) studies on proteins. These new tools constitute an essential enabling technology to determine the structures and functional dynamics for a wide range of soluble and membrane-bound proteins and their complexes. As such, these tools should have an immediate and significant impact on SDSL studies of proteins here at Vanderbilt and elsewhere, and we intend to make all software and computational protocols developed in this program project freely available to colleagues at other institutions. The specific aims of this program project are: 1) develop molecular dynamics and molecular modeling protocols to enable us to directly correlate EPR measurements of spin label mobility and accessibility with protein structure and structural fluctuations; 2) develop simulation tools and protocols so that EPR measurements of inter-probe distances can be related directly to protein structure; 3) develop a general computational protocol so that information obtained in Specific Aims 1 and 2 can be used to build and refine structural models; and 4) apply the tools and strategies developed in Aims 1-3 to SDSL data obtained for T4 lysozyme, a model protein of known structure, for CDB3, a more complex erythrocyte membrane protein that is to date only partially characterized structurally, and for amyloid precursor protein peptides that are not well characterized structurally at this time. These specific protein spectroscopic studies will provide the requisite experimental data necessary to develop and test the computational protocols. This program includes investigators with extensive eperience in EPR spectroscopy and SDSL studies of proteins (Beth and Hustedt), membrane protein structure-function studies (Beth, Lybrand, and Sanders), and use of molecular modeling and molecular simulation tools to refine three-dimensional protein structures from distance data obtained from spectroscopic and other biophysical studies (Lybrand and Smith).

该计划项目的长期目标是开发计算工具，这些工具可用于从蛋白质上的定向旋转标记（SDSL）研究中提取准确的结构和动力学信息。这些新工具构成了一种必不可少的技术，可以确定各种可溶性和膜结合蛋白及其复合物的结构和功能动力学。因此，这些工具应该对范德比尔特和其他地方的蛋白质研究的SDSL研究产生直接和重大的影响，我们打算使该计划项目中开发的所有软件和计算协议在其他机构的同事中免费使用。该程序项目的具体目的是：1）开发分子动力学和分子建模方案，使我们能够直接将自旋标签迁移率和可访问性的EPR测量与蛋白质结构和结构波动相关联； 2）开发模拟工具和协议，以便对探针间距离的EPR测量可以直接与蛋白质结构有关； 3）制定一般的计算协议，以便可以使用特定目标1和2中获得的信息来构建和完善结构模型； 4）将目标1-3中开发的工具和策略应用于针对T4溶菌酶（已知结构的模型蛋白）CDB3获得的SDSL数据，CDB3是一种更复杂的红细胞膜蛋白，迄今为止仅部分在结构上部分地表征了部分特征，并且对于氧化肌体蛋白蛋白质蛋白质的肽在结构上没有很好地表征。这些特定的蛋白质光谱研究将提供开发和测试计算方案所需的必要实验数据。该计划包括在EPR光谱法和蛋白质的SDSL研究（Beth和Hustedt），膜蛋白结构功能研究（Beth，Lybrand和Sanders）中具有广泛敏捷性的研究人员，以及分子建模和分子模拟的使用，用于从远距离数据中获得的三维蛋白质结构，并从spectrrasmscic and Smith smith smith smith shiph smith shiper shiper shiper shiper shime of。