X-RAY AND NEUTRON SCATTERING STUDIES OF COLLECTIVE MOTIONS IN PROTEINS

蛋白质集体运动的 X 射线和中子散射研究

• 批准号: 7954900
• 负责人: LEE MAKOWSKI
• 金额: $ 2.72万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954900
• 关键词: Accounting; Affect; Behavior; Biophysics; Computer Retrieval of Information on Scientific Projects Database; Crystallography; Data; Elements; Environment; Exhibits; Funding; Glutamate Receptor; Grant; HIV Protease; Hemoglobin; Institution; Ligation; Link; Measures; Molecular Structure; Motion; Muramidase; Mutation; Neutrons; Pathway interactions; Pattern; Process; Proteins; Protocols documentation; Relative (related person); Reproducibility; Research; Research Personnel; Resources; Roentgen Rays; Solutions; Source; Structure; Structure-Activity Relationship; United States National Institutes of Health; novel strategies; protein structure function; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Proteins in solution exhibit an ensemble of interconverting structures accessible to one another through low energy pathways. The breadth and form of this ensemble defines the dynamic behavior of the protein. It is increasingly clear that protein structure-function relationships must account for this native ensemble and its associated dynamics. Characterization of the native ensembles of a protein represents a significant technical challenge, and although recent advances in NMR, x-ray crystallography, and computational approaches have contributed substantially, new approaches are badly needed. We have demonstrated that wide-angle x-ray solution scattering (WAXS) can be used to estimate the breadth of the native ensemble of proteins in solution. Recent improvements in protocols for collecting and processing WAXS data have resulted in a substantial improvement in accuracy and reproducibility. These advances provide the opportunity to accurately measure second order effects in the scattering patterns making possible their interpretation in terms of changes in the form and breadth of the ensemble of macromolecular structures in solution. Here we will use WAXS to characterize the native state ensemble of a number of representative proteins and study how changes in ligation state, environment and functionally relevant mutations affect the ensemble. Proteins to be studied include hemoglobin, HIV protease, glutamate receptor and lysozyme. We will show that WAXS can be used to estimate the amplitudes of a protein's normal modes and thereby act as an important new tool for establishing a direct link between a protein's environment or ligation state and the relative motions of specific structural elements within it.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 溶液中的蛋白质表现出相互转换结构的集合，通过低能量途径相互接近。 这个集合的宽度和形式定义了蛋白质的动态行为。 越来越清楚的是，蛋白质的结构-功能关系必须解释这种天然的整体及其相关的动力学。 蛋白质天然集合体的表征是一项重大的技术挑战，尽管核磁共振、X射线晶体学和计算方法的最新进展做出了重大贡献，但仍然迫切需要新的方法。 我们已经证明，广角X-射线溶液散射（WAXS）可以用来估计的广度的天然系综的蛋白质在溶液中。最近在收集和处理WAXS数据的协议中的改进导致了准确性和再现性的大幅提高。 这些进展提供了机会，以准确地测量二阶效应的散射图案，使其解释在溶液中的大分子结构的合奏的形式和宽度的变化。 在这里，我们将使用WAXS来表征一些代表性蛋白质的天然状态集合，并研究连接状态，环境和功能相关突变的变化如何影响集合。待研究的蛋白质包括血红蛋白、HIV蛋白酶、谷氨酸受体和溶菌酶。我们将表明，WAXS可用于估计蛋白质的正常模式的振幅，从而作为一个重要的新工具，用于建立蛋白质的环境或连接状态和特定结构元素的相对运动之间的直接联系。