WAXS AS A PROBE FOR THE STUDY OF PROTEIN STRUCTURE, DYNAMICS AND FUNCTION

蜡作为研究蛋白质结构、动力学和功能的探针

• 批准号: 8168619
• 负责人: LEE MAKOWSKI
• 金额: $ 2.2万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168619
• 关键词: Behavior; Binding; Catalysis; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Data Collection; Event; Funding; Goals; Grant; HIV Protease; Hemoglobin; Institution; Ligand Binding; Ligands; Measurement; Measures; Membrane Proteins; Molecular; Motion; Mutation; Protein Dynamics; Proteins; Research; Research Personnel; Resources; Screening procedure; Signaling Protein; Solutions; Source; Structure; Temperature; United States National Institutes of Health; adenylate kinase; protein function; protein structure; success; 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. Wide-angle x-ray solution scattering (WAXS) has significant potential for characterizing the structure, dynamics and function of proteins without the need for crystallization (Fischetti et al., 2003). We have carried out a theoretical analysis of the information content of WAXS and quantitated its capabilities for determining the secondary and tertiary structure of a protein (Makowski et al., 2008a). Using WAXS from proteins at different concentrations or temperatures, we demonstrated that WAXS contains substantial information about the structural fluctuations within an ensemble of proteins (Makowski et al., 2008b). We have further shown that the amplitudes of normal modes of motion of a protein can be estimated using WAXS (Park and Makowski, 2008). We have carried out the first measurement of the anomalous signal from proteins in solution, and demonstrated that it can be used to measure interatomic distances within proteins (Fischetti et al., 2008), including membrane proteins (Makowski et al., 2008c). Its power for measurement of small structural changes makes it an excellent tool for screening of functional binding events (Rodi et al., 2007). The goal of the present request is to continue data collection on a number of projects that will expand and develop these recent successes. In particular, we will carry out several related projects that will utilize WAXS for the study of the structure and dynamics of proteins, including: (i) A detailed analysis of the effect of mutations on the dynamic behavior of HIV protease; (ii) Estimation of the breadth of molecular motion during catalysis of HIV protease; (iii) Analysis of the effect of ligand state and mutations on the dynamic behavior of hemoglobin; (iv) analysis of the structural changes that occur during ligand binding and catalytic activity of adenylate kinase.

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。 广角X射线溶液散射（WAXS）具有在不需要结晶的情况下表征蛋白质的结构、动力学和功能的显著潜力（Fischetti等人，2003年）。 我们已经对WAXS的信息内容进行了理论分析，并量化了其确定蛋白质二级和三级结构的能力（Makowski等人，2008年a）。 使用来自不同浓度或温度下的蛋白质的WAXS，我们证明WAXS包含关于蛋白质系综内的结构波动的大量信息（Makowski等人，2008年b）。 我们进一步表明，蛋白质的正常运动模式的振幅可以使用WAXS来估计（Park和Makowski，2008）。 我们已经对溶液中蛋白质的异常信号进行了首次测量，并证明其可用于测量蛋白质内的原子间距离（Fischetti et al.，2008），包括膜蛋白（Makowski et al.，2008年c）。 其测量小结构变化的能力使其成为筛选功能性结合事件的极好工具（Rodi等人，2007年）。 本请求的目的是继续收集关于若干项目的数据，这些项目将扩大和发展最近取得的成功。 特别是，我们将开展几个相关项目，将利用WAXS研究蛋白质的结构和动力学，包括：（i）突变对HIV蛋白酶动力学行为的影响的详细分析;（ii）HIV蛋白酶催化过程中分子运动宽度的估计;（iii）配体状态和突变对血红蛋白动力学行为的影响的分析;（iv）分析腺苷酸激酶的配体结合和催化活性期间发生的结构变化。