Mapping the alcohol site of a neuronal potassium channel

绘制神经元钾通道的酒精位点

• 批准号: 7760976
• 负责人: MANUEL L COVARRUBIAS
• 金额: $ 24.17万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7760976
• 关键词: Acute; Address; Alanine; Alcohol dependence; Alcoholic Intoxication; Alcohols; Amino Acid Substitution; Anesthetics; Architecture; Attention; Binding; Binding Sites; Brain; C-terminal; Chemosensitization; Coupling; Development; Drosophila genus; Electrophysiology (science); Engineering; Ethanol; Exhibits; Funding; General anesthetic drugs; Genetic Engineering; Goals; Hydrogen Bonding; Ion Channel; Ion Channel Protein; Kinetics; Laboratories; Length; Light; Lipid Bilayers; Maps; Methods; Modeling; Molecular; Movement; Mutagenesis; National Institute on Alcohol Abuse and Alcoholism; Neurons; Organism; Pharmaceutical Preparations; Physiological; Positioning Attribute; Potassium Channel; Probability; Property; Proteins; Research Personnel; Resistance; Role; Scanning; Series; Site; Solutions; Structure; Study Subject; System; Tail; Testing; Therapeutic; Transplantation; Voltage-Gated Potassium Channel; Work; alcohol effect; alcohol sensitivity; base; insight; interest; member; mutant; patch clamp; programs; protein purification; protein structure; reconstitution; response; structural biology; synthetic peptide; three dimensional structure

DESCRIPTION (provided by applicant): The long-term goal of this project is to determine the structural basis of the functional modulation of brain ion channels by alcohol. The subject of this study is the neuronal Shaw2 potassium channel, which is directly inhibited by pharmacologically relevant concentrations of ethanol and other short-chain members of the homologous series of 1-alkanols. Guided by previous results from this study, the central hypothesis of the project states that the activation gate of the Shaw2 channel contributes an amphipathic protein-protein interface that constitutes the alcohol binding site. This project focuses on investigating the activation gate as the locus of the interactions that control the alcohol-induced responses of the Shaw2 channel. The specific aims of the project are: 1) To investigate the specific structural determinants of alcohol binding in the S4-S5 loop of the Shaw2 potassium channel. 2) To investigate the structural features that govern the alcohol-induced responses of the Shaw2 potassium channel in the C-terminal section of the S6 segment. 3) To develop the prokaryotic KvAP channel as a model to explore the achitecture of the alcohol binding site in the Shaw2 potassium channel. The first two aims systematically combine recombinant DNA technology, electrophysiology and biophysical analyses to map the alcohol site in the Shaw2 channel. In light of the recently solved crystal structure of the KvAP channel, the last aim seeks the potential application of structural biology to determine the achitecture of an engineered alcohol site. A detailed map of the physiologically relevant amphipathic interfaces that bind alcohol in ion channels is the first step toward understanding acute alcohol intoxication at the atomic level and targeting these sites for therapeutic applications.

描述（由申请人提供）：该项目的长期目标是确定通过酒精对脑离子通道的功能调节的结构基础。这项研究的主题是神经元SHAW2钾通道，该通道被药理学相关的乙醇和同源系列1-烷醇的其他短链成员直接抑制。在这项研究的先前结果的指导下，该项目的中心假设指出，Shaw2通道的激活门贡献了构成酒精结合位点的两亲性蛋白质蛋白界面。该项目的重点是研究激活门，作为控制Shaw2通道的酒精引起的反应的相互作用的源。该项目的具体目的是：1）研究shaw2钾通道S4-S5环中酒精结合的特定结构决定因素。 2）研究在S6节段的C末端部分中酗酒诱导的shaw2钾通道反应的结构特征。 3）开发原核KVAP通道作为探索Shaw2钾通道中酒精结合位点的成就的模型。前两个目的系统地结合了重组DNA技术，电生理学和生物物理分析，以绘制Shaw2通道中的酒精位点。鉴于KVAP通道的最近解决的晶体结构，最后一个目的寻求结构生物学的潜在应用来确定工程酒精位点的实现。在离子通道中结合酒精的生理相关两亲界面的详细图是理解原子水平急性酒精中毒的第一步，并将这些位点靶向用于治疗应用。