NMR STUDIES OF BIOMOLECULAR STRUCTURE, FUNCTION, AND DYNAMICS

工作总结：本项目的总体目标涉及使用 核磁共振波谱在生物表征中的应用 大分子及其与环境化合物的相互作用 担忧。环境因子形成蛋白质加合物的研究进展 涉及复杂的化学/生化/结构相互作用， 充其量，也就是不完全理解。到目前为止，这项工作涉及几个 具有环境和药理价值的药物，包括 溴乙酸酯，一种羧甲基化试剂，以及阿司匹林。二维图 质子-碳HMQC实验可以分辨加合物的共振 与单个残基反应形成的。解决问题的几种方法 对这些共振峰的归属进行了评估。这些研究已经 涉及两种目标蛋白质：泛素--一种小而稳定的蛋白质 缺乏活性部位，以及最近的血红蛋白。尽管 阿司匹林乙酰化的主要生理靶点被认为是 前列腺素合成酶，阿司匹林已知可以乙酰化许多其他 蛋白质，例如白蛋白和血红蛋白，以及这种相互作用可能会导致 具有广泛的药理和毒理作用。这项研究 最近根据与Dr。 理查德·拉博特卡一直在调查各种能够 干扰镰状血红蛋白的聚集。阿司匹林是其中之一 为此目的而研究的最早的药物，并已知可修饰 DPG结合口袋中的残留物。拉博特卡博士最近研制出 几种用于此目的的改良试剂，我们正在研究血红蛋白 由这些代理进行的修改是一个协作项目。相对于 阿司匹林和其他已为此目的进行评估的药物。 Labotka的化合物被预测具有更好的传输性能 而且应该不会对正常的细胞新陈代谢造成太大的干扰。一个 与Linda Luck博士(克拉克森大学)合作研究项目 在大肠杆菌上，周质葡萄糖-半乳糖受体仍在继续。一个 基于观察到的动态频移行为的手稿 13C，1-2H]葡萄糖与受体的络合最近完成，并且 对氟色氨酸标记受体的进一步研究正在进行中。

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.