Measuring static and dynamic electric fields in proteins

测量蛋白质中的静态和动态电场

• 批准号: 7807133
• 负责人: Nicholas Mark Levinson
• 金额: $ 4.76万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7807133
• 关键词: Active Sites; Affect; Aldehyde Reductase; Area; Behavior; Binding; Biochemical; Biological; Biological Process; Catalysis; Development; Diabetes Mellitus; Disease; Electrostatics; Environment; Enzymes; Evolution; GTP-Binding Proteins; GTPase-Activating Proteins; Guanosine Triphosphate Phosphohydrolases; Human; Lead; Malignant Neoplasms; Measurement; Measures; Methods; Molecular Conformation; Monomeric GTP-Binding Proteins; Mutate; Mutation; Nitriles; Pharmaceutical Preparations; Play; Process; Property; Protein Analysis; Protein Dynamics; Protein Region; Proteins; Reporter; Research; Role; Site; Specificity; Spectrum Analysis; System; Techniques; Therapeutic; Work; biological systems; comparative; design; electric field; improved; inhibitor/antagonist; insight; interest; macromolecule; physical property; protein function; protein structure function; public health relevance; research study

DESCRIPTION (provided by applicant): The long-term scientific objectives of this application are to pursue an area of research that moves beyond a traditional structural and biochemical analysis of proteins to a more quantitative understanding of the energetics that govern protein function. Electric fields are an integral component of the energy landscapes that govern protein structure and function but very few experimental measurements of electric fields in proteins have been made. A spectroscopic technique called VSE Spectroscopy, developed by Steve Boxer's research group, will be used to perform a systematic characterization of the electric fields in two biological systems. Specific aim 1 involves comparing the electric fields in aldose and aldehyde reductase, two enzymes implicated in diabetes. This will reveal how the active sites of these closely related enzymes are electrostatically tuned to different functions, and how these differences could be used to design more potent and more specific inhibitors of aldose reductase. Specific Aim 2 involves studying the small GTPase Ras, which is mutated in a large number of human cancers. Since Ras is a highly dynamic protein, this work will enhance our understanding of how electrostatics is influenced by protein dynamics, a question about which relatively little is known. VSE Spectroscopy requires the incorporation of a nitrile group into the protein of interest to serve as an experimental reporter of electric field. Two methods have been previously developed for the incorporation of nitriles into proteins and these methods will be used to prepare proteins with probes at different sites, allowing the electrostatics to be measured over large regions of the proteins of interest. Since the Boxer lab pioneered the technique of VSE Spectroscopyit will provide an ideal environment for performing these experiments. PUBLIC HEALTH RELEVANCE: Many diseases are caused by relatively subtle perturbations to specific biological processes- for instance, cancer involves the acquisition of mutations that alter the properties of certain proteins. A comprehensive understanding of the physical properties of such proteins will reveal the origin of the aberrant behavior that arises in disease and allow us to exploit these properties for therapeutic benefit. The work proposed here with aldose reductase and Ras will improve our understanding of the involvement of these proteins in disease and lead to new ways to intervene in the disease process.

描述（由申请人提供）：本应用的长期科学目标是追求研究领域，该领域超越了对蛋白质的传统结构和生化分析，以对控制蛋白质功能的能量学有更定量的理解。电场是控制蛋白质结构和功能的能量景观的组成部分，但对蛋白质中电场的实验测量很少。由史蒂夫·拳击手研究小组开发的一种称为VSE光谱的光谱技术将用于在两个生物系统中对电场进行系统表征。具体目标1涉及比较醛和醛还原酶中的电场，两种与糖尿病有关的酶。这将揭示这些密切相关的酶的活性位点如何静电调节到不同的功能，以及如何使用这些差异来设计醛糖还原酶的更有效和更特异性的抑制剂。具体目标2涉及研究小型GTPase RA，该RA在大量的人类癌症中被突变。由于RAS是一种高度动态的蛋白质，因此这项工作将增强我们对静电如何受蛋白质动力学影响的理解，这是一个相对较少的问题。 VSE光谱法要求将氮基团掺入感兴趣的蛋白质中，以作为电场的实验记者。先前已经开发出两种方法将硝酸盐掺入蛋白质中，这些方法将用于制备在不同位点使用探针的蛋白质，从而使静电可以在感兴趣的蛋白质的大区域进行测量。由于Boxer Lab开创了VSE Spectroscopyit的技术，将为执行这些实验提供理想的环境。公共卫生相关性：许多疾病是由对特定生物过程的相对微妙的扰动引起的 - 例如，癌症涉及获取改变某些蛋白质特性的突变。对此类蛋白质的物理特性的全面了解将揭示出在疾病中产生的异常行为的起源，并使我们能够利用这些特性为治疗益处。在这里提出的用醛糖还原酶和RAS提出的工作将提高我们对这些蛋白质参与疾病的参与的理解，并带来新的干预疾病过程的方法。