The molecular basis of alcohol's actions.

酒精作用的分子基础。

• 批准号: 6848351
• 负责人: DAVID NIGEL JONES
• 金额: $ 29.31万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6848351
• 关键词: GABA receptor; NMDA receptors; X ray crystallography; binding proteins; binding sites; butanols; chemical stability; ethanol; fluorescence spectrometry; intermolecular interaction; ligands; nuclear magnetic resonance spectroscopy; point mutation; propanols; protein engineering; protein structure function; site directed mutagenesis

DESCRIPTION (provided by applicant): Objectives and Health Related Issues. The intoxicating effects associated with exposure to ethanol have been linked to changes in the activities of several neurotransmitter receptors that are ligand-gated ion-channels. Acute exposure of the gamma-amino-butyric acid (GABA) and N-methyl D-aspartate (NMDA) receptors to ethanol is directly implicated in the development of fetal alcohol syndrome and also in alcohol toxicity and alcohol dependency in the adult. There is increasing evidence that ethanol binds to specific sites on these receptors and induces a conformational change that modifies their activity. Characterization of alcohol-binding sites in ethanol sensitive proteins would provide potential targets for the development of pharmacological agents to control alcohol intoxication and alcohol dependency. At present there is no direct structural information available about the nature of potential binding sites of these important ethanol-sensitive proteins because of the inherent difficulties in studying integral membrane proteins. LUSH is a novel alcohol-binding protein from fruit flies that recognizes ethanol, n-propanol and n-butanol. We have recently solved the structure of LUSH in the complex with ethanol. The long-term goal of this proposal is to characterize the molecular nature of this binding site in detail in order to define the molecular basis for alcohol-binding specificity. The structure of LUSH bound to a series of alcohols will be solved using X-ray crystallographic methods to reveal a molecular picture of alcohol specificity in a nonenzymatic protein. The effect of different alcohols on binding affinity and protein stability will be analyzed using biophysical and spectroscopic methods, and the differences correlated with changes to the protein structure in solution. The role of specific amino acids in alcohol binding and protein function will be tested using site directed mutagenesis to engineer proteins with modified ligand-binding properties. The ultimate goal is to develop a model for specific alcohol-binding sites in alcohol-sensitive proteins that will aid in an understanding of the molecular basis of alcohols actions in causing intoxication and toxicity.

描述（由申请人提供）：目标和与健康有关的问题。与接触乙醇有关的醉人作用与配体门控离子通道的几种神经递质受体的活性变化有关。 γ-氨基丁酸（GABA）和N-甲基D-天冬氨酸（NMDA）受体急性暴露于乙醇直接与胎儿酒精综合征的发展以及成人的酒精毒性和酒精依赖性有关。越来越多的证据表明，乙醇与这些受体上的特定位点结合，并诱导构象变化，从而改变其活性。乙醇敏感蛋白中酒精结合位点的表征将为制定药理剂的开发提供潜在的靶标，以控制酒精中毒和酒精依赖性。目前，由于研究整合膜蛋白的固有困难，目前尚无有关这些重要乙醇敏感蛋白潜在结合位点的性质的直接结构信息。郁郁葱葱的是一种新型的酒精结合蛋白，从水果蝇中识别出乙醇，N-丙醇和N-丁醇。最近，我们解决了与乙醇综合体中郁郁葱葱的结构。该建议的长期目标是详细表征该结合位点的分子性质，以定义酒精结合特异性的分子基础。将使用X射线晶体学方法来解决与一系列酒精结合的郁郁葱葱的结构，以揭示非酶蛋白中酒精特异性的分子图片。不同醇对结合亲和力和蛋白质稳定性的影响将使用生物物理和光谱方法分析，并且差异与溶液中蛋白质结构的变化有关。特异性氨基酸在酒精结合和蛋白质功能中的作用将使用定向诱变对具有改良配体结合特性的工程蛋白的定向诱变。最终目标是为酒精敏感蛋白中特定的酒精结合位点开发模型，这将有助于理解酒精的分子基础，从而引起中毒和毒性。