PROTEIN MOTIONS IN RECOGNITION, REGULATION AND CATALYSIS

识别、调节和催化中的蛋白质运动

• 批准号: 7228121
• 负责人: ARTHUR G PALMER
• 金额: $ 31.88万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7228121
• 关键词: Active Sites; Address; Agonist; Amino Acids; Aprotinin; Binding; Biological; Biological Models; Biological Process; Breathing; C-terminal; Carboxy-Lyases; Catalysis; Cellular Structures; Chemicals; Closure; Complex; Computing Methodologies; Condition; Core Protein; Coupled; Coupling; Development; Disease; Drug Addiction; Enzyme Inhibition; Enzymes; Equilibrium; Event; Glutamate Receptor; Goals; Hereditary Disease; Homologous Protein; Investigation; Isomerase; Kinetics; Laboratories; Lead; Ligand Binding; Ligand Binding Domain; Ligands; Magnetism; Malignant Neoplasms; Measurement; Measures; Mechanics; Metabolism; Methods; Microscopic; Modeling; Modification; Molecular; Molecular Conformation; Monitor; Motion; Multienzyme Complexes; Mutation; N-terminal; NMR Spectroscopy; Nuclear; Numbers; Ornithine Decarboxylase; Pathology; Pathology, Other; Phase; Population; Process; Protein Dynamics; Proteins; Range; Rate; Reaction; Receptor Activation; Regulation; Relative (related person); Relaxation; Relaxation Techniques; Research; Research Personnel; Resolution; Role; Scheme; Side; Site; Site-Directed Mutagenesis; Solutions; Structure; System; Techniques; Testing; Thermodynamics; Time; Ubiquitin; Uridine Monophosphate; base; conformer; desensitization; dimer; gain of function; in vitro Model; inhibitor/antagonist; inorganic phosphate; insight; interest; intermolecular interaction; ionization; method development; millisecond; molecular recognition; mutant; neurotransmission; novel; orotidylic acid; programs; protein activation; protein folding; protein function; protein protein interaction; quantum; research study; response; solid state; technique development; villin

DESCRIPTION (provided by applicant): Recent developments, including those from the applicant laboratory, have opened new opportunities for investigation of dynamic processes on mu s-ms time scales using NMR spin relaxation measurements. Motions on these time scales reflect large-amplitude loop motions, relative motions between domains, collective "breathing" of protein cores, ligand-binding or oligomerization reactions, and overall folding-unfolding events. Such processes may be closely coupled, and in some instances rate-limiting, to biological functions such as molecular recognition, transitions along the catalytic cycle of enzymes, and inhibition or activation of proteins through intra-or inter-molecular protein-protein interactions. Mutations that perturb dynamical processes and conformational equilibria can be associated with significant pathology, including loss or gain of function and misfolding. The existence of large amplitude intra-molecular conformational changes in proteins have been inferred from comparison of equilibrium structures of a given protein in different crystal forms, a given protein in free and ligand-bound states, or sets of homologous proteins. However, only solution NMR spectroscopy can confirm the occurrence and determine the kinetic rates in the solution state of dynamic processes, at equilibrium and with atomic site resolution, in the absence of influences from intermolecular interactions in the solid state, and without potential complications introduced by non-native modifications necessary for other solution-state spectroscopic techniques. The proposed research has four primary objectives: (1) elucidation of the folding mechanism and description of the unfolded-state ensemble for the villin headpiece domain HP67; (2) identification of the mechanistic basis for coupling between agonist or antagonist binding and function of the ionotropic glutamate receptor GluR2 S1S2 ligand-binding domain; (3) assessment of the role of conformational mobility in catalysis by ornithine decarboxylase; and (4) development of novel experimental and theoretical methods for characterizing protein dynamics on mu s-ms time scales. Successful completion of these goals will enable applications to a wide range of protein systems of biological interest. Time-dependent structural changes underlie the normal function of proteins, and misfunction in genetic diseases, cancer, and other pathologies. The proposed research will measure these changes for three model protein systems involved in cellular structure, nerve transmission, and basic metabolism.

描述（由申请人提供）：最近的发展，包括申请人实验室的发展，为使用NMR旋转放松测量值开辟了新的机会，以调查MU S-MS时间尺度上的动态过程。这些时间尺度上的运动反映了巨大的振幅回路运动，域之间的相对运动，蛋白质核心的集体“呼吸”，配体结合或寡数反应以及整体折叠 - 不折叠事件。此类过程可以紧密耦合，在某些情况下，限制速率，与生物学功能，例如分子识别，沿酶的催化循环过渡以及通过内部或分子间蛋白质蛋白质相互作用的蛋白质抑制或激活。扰动动力学过程和构象平衡的突变可能与重要的病理学有关，包括功能的丧失或折叠率。通过比较给定蛋白的平衡结构以不同的晶体形式的平衡结构，在自由和配体结合的状态或一组同源蛋白质中的给定蛋白质的比较，已经推断出蛋白质中的大分子内构象变化的存在。但是，只有溶液NMR光谱可以确认发生的发生，并确定动态过程的溶液状态，平衡和原子位点分辨率的溶液状态，如果没有固态中分子间相互作用的影响，并且没有潜在的并发症，而没有由其他溶液溶液溶液STATE STATE STATE STATESPATESPOCTROCCOCICECES介绍的潜在并发症。拟议的研究具有四个主要目标：（1）阐明了Villin头把面域HP67的折叠机制和描述； （2）鉴定激动剂或拮抗剂结合和拮抗剂谷氨酸受体Glur2 S1S2配体结合结构域之间耦合的机理基础； （3）评估构象迁移率在鸟嘌呤脱羧酶催化中的作用； （4）开发用于表征MU S-MS时间尺度上蛋白质动力学的新型实验和理论方法。这些目标的成功完成将使应用于生物学关注的广泛蛋白质系统。时间依赖性的结构变化是蛋白质的正常功能，以及遗传疾病，癌症和其他病理学的功能错误。拟议的研究将针对参与细胞结构，神经传播和基本代谢的三种模型蛋白系统的这些变化衡量这些变化。