NMR STUDIES OF BIOMOLECULAR STRUCTURE, FUNCTION, AND DYNAMICS

生物分子结构、功能和动力学的核磁共振研究

• 批准号: 6162236
• 负责人: R E LONDON
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6162236
• 关键词: aspirin; blood /lymphatic pharmacology; chemical structure function; drug metabolism; environmental toxicology; hemoglobin; intermolecular interaction; microorganism culture; molecular dynamics; nuclear magnetic resonance spectroscopy; toxin; ubiquitin

Summary of Work: The overall aim of this project involves the use of nuclear magnetic resonance (NMR) spectroscopy to characterize biological macromolecules and their interaction with compounds of environmental concern. The formation of protein adducts by environmental agents involves complex chemical/biochemical/structural interactions which are, at best, incompletely understood. Work to date has involved several agents of environmental and pharmacological interest, including bromoacetate, a carboxymethylating agent, and aspirin. Two dimensional proton-carbon HMQC experiments allow resolution of the adduct resonances formed from reaction with individual residues. Several approaches to the assignment of these resonances have been evaluated. These studies have involved two target proteins: ubiquitin - a small, stable protein lacking an active site, and, more recently, hemoglobin. Although the primary physiological target of acetylation by aspirin is believed to be prostaglandin synthase, aspirin is known to acetylate many other proteins, e.g. albumin and hemoglobin, and such interactions can result in a broad range of pharmacological and toxicological effects. The study of hemoglobin was recently initiated based on discussions with Dr. Richard Labotka who has been investigating various agents capable of interfering with the aggregation of sickle hemoglobin. Aspirin was one of the earliest agents studied for this purpose, and is known to modify residues in the DPG binding pocket. Dr. Labotka has recently developed several improved agents for this purpose, and we are studying hemoglobin modifcation by these agents as a collaborative project. Relative to aspirin and other agents which have been evaluated for this purpose, Dr. Labotka's compounds are predicted to have improved transport properties and should be less perturbing to normal cellular metabolism. A collaborative research project with Dr. Linda Luck (Clarkson University) on the E. coli periplasmic glucose-galactose receptor has continued. A manuscript based on the dynamic frequency shift behavior observed for [1- 13C,1-2H]glucose complexed with the receptor was recently completed, and further studies of fluorotryptophan labeled receptor are in progress.

工作总结：本项目的总体目标涉及使用 核磁共振（NMR）光谱学来表征生物 大分子及其与环境化合物的相互作用 关心环境因子对蛋白质加合物的形成 涉及复杂的化学/生物化学/结构相互作用， 最多也就是不完全理解 迄今为止， 具有环境和药理学意义的物质，包括 溴乙酸盐、羧甲基化剂和阿司匹林。 二维 质子碳HMQC实验允许加合物共振的分辨率 由单个残基反应形成。 几种方法 已经评估了这些共振的分配。 这些研究 涉及两种靶蛋白：泛素-一种小而稳定的蛋白 缺乏活性部位最近还有血红蛋白 虽然 阿司匹林乙酰化的主要生理靶点被认为是 前列腺素合酶，已知阿司匹林乙酰化许多其他的前列腺素合酶。 蛋白质，例如白蛋白和血红蛋白，并且这种相互作用可导致 具有广泛的药理学和毒理学作用。 研究 血红蛋白是最近发起的讨论的基础上，博士。 理查德·拉博特卡一直在调查各种特工， 干扰镰状血红蛋白的聚集。 阿司匹林是其中之一 最早为此目的研究的代理人，并已知修改 DPG结合口袋中的残基。 拉博特卡博士最近开发了 几种改进的药物，我们正在研究血红蛋白 由这些代理人作为一个合作项目进行修改。 相对于 阿司匹林和其他药物已被评估用于这一目的，博士。 据预测，Labotka的化合物具有更好的传输性能 并且对正常细胞代谢的干扰较小。 一 与琳达勒克博士（克拉克森大学）的合作研究项目 在E。大肠杆菌周质葡萄糖-半乳糖受体的表达仍在继续。 一 手稿的基础上观察到的动态频移行为[1- 13 C，1- 2 H]葡萄糖与受体的复合最近完成， 对荧光色氨酸标记受体的进一步研究正在进行中。