NMDA receptors with restricted mobility of the ligand binding domain

配体结合域移动性受限的 NMDA 受体

基本信息

批准号：
7450118
负责人：
Gabriela K Popescu
金额：
$ 7.93万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7450118
关键词：
6,7-dichloroquinoxaline-2,3-dione Abbreviations Acids Acute Agonist Alkanesulfonates Alzheimer&apos s Disease Arts Behavior Binding Binding Sites Biological Brain Cations Cells Chronic Cleaved cell Closure Cognition Disorders Condition Cross-Linking Reagents Cycloleucine Cysteine Depth Development Disulfides Dithionitrobenzoic Acid Dithiothreitol Drug Delivery Systems Drug effect disorder Engineering Epilepsy Event Excitatory Synapse Foundations Glutamate Agonist Glutamate Receptor Glutamates Glycine Hydrogen Peroxide Investigation Ion Channel Kinetics Knowledge Length Ligand Binding Ligand Binding Domain Ligands Link Lobe Measurable Membrane Mental Retardation Mental disorders Methane Molecular Molecular Conformation Motion N-Methyl-D-Aspartate Receptors NR1 gene Nerve Degeneration Neurons Neurotransmitter Receptor Neurotransmitters Oxidants Pathway interactions Phenanthrolines Physiological Physiology Proteins Public Health Rate Reaction Reagent Receptor Activation Reducing Agents Role Schizophrenia Site Stroke Sulfhydryl Compounds Testing Therapeutic Time addiction chronic pain crosslink cyclopropanecarboxylic acid design extracellular human NR1 protein insight mutant novel therapeutics prevent receptor receptor function research study single molecule

项目摘要

DESCRIPTION (provided by applicant): NMDA receptors are membrane-bound neurotransmitter receptors with critical roles in brain physiology. They are principal drug targets for Alzheimer's disease and stroke, mental retardation, and also addiction, chronic pain, schizophrenia and epilepsy. NMDA receptors are glutamate-activated, excitatory ion-channels. Their activation reaction is initiated when the neurotransmitter glutamate binds to extracellular, ligand-binding domains (LBDs) and culminates with the opening of a membrane-embedded cation-selective pore. The molecular events that make up the activation reaction of NMDA receptors remain unknown, despite their fundamental role in controlling receptor function. Kinetic studies of single-receptor activity have established that after binding agonists, NMDA receptors cycle with measurable rates between several closed-pore and open-pore conformations. Structural studies of the isolated LBDs have shown that ligands bind deep inside a crevice between two mobile lobes and have put forth the hypothesis that ligand-induced closure of the LBD is one of the required steps along the NMDA receptor activation pathway. This application proposes experiments that will directly and specifically test this hypothesis by examining the activity of NMDA receptors with restricted mobility of the LBDs. Pairs of cysteine residues will be introduced at the tips of the LBDs lobes as a means to lock these at defined distances with respect to one another. The engineered receptors will be examined by kinetic analyses of their macroscopic and single-channel behaviors in the presence and absence of oxidizing agents, and following treatment with crosslinking reagents of various lengths. The results will provide basic information regarding the identity of the molecular motions associated with NMDA receptor activation and will afford valuable insight regarding possible mechanisms of drug action at this receptor. This knowledge will form the conceptual foundation necessary to rationally control NMDA receptor activities as a therapeutic strategy for stroke, addiction, and mental illness. PUBLIC HEALTH RELEVANCE: This project will provide fundamental information regarding the mechanism by which NMDA receptors become active. This information is currently lacking and is needed for the rational design of novel therapeutic approaches for acute and chronic neurodegeneration (stroke, Alzheimer's disease), addiction, chronic pain, cognitive disorders and mental illness.

描述（由申请人提供）：NMDA 受体是膜结合神经递质受体，在脑生理学中具有关键作用。它们是治疗阿尔茨海默病、中风、智力低下、成瘾、慢性疼痛、精神分裂症和癫痫的主要药物靶标。 NMDA 受体是谷氨酸激活的兴奋性离子通道。当神经递质谷氨酸与细胞外配体结合域 (LBD) 结合时，它们的激活反应就会启动，并随着膜嵌入的阳离子选择性孔的打开而达到顶峰。构成 NMDA 受体激活反应的分子事件仍然未知，尽管它们在控制受体功能方面发挥着基本作用。单受体活性的动力学研究表明，在结合激动剂后，NMDA 受体在几种闭孔和开孔构象之间以可测量的速率循环。对分离的 LBD 的结构研究表明，配体在两个移动叶之间的缝隙深处结合，并提出了这样的假设：配体诱导的 LBD 闭合是 NMDA 受体激活途径所需的步骤之一。本申请提出的实验将通过检查限制 LBD 移动性的 NMDA 受体的活性来直接、具体地检验这一假设。成对的半胱氨酸残基将被引入到 LBD 叶的尖端，作为将这些残基彼此锁定在规定距离处的一种手段。将通过在存在和不存在氧化剂的情况下以及在用不同长度的交联剂处理后对其宏观和单通道行为进行动力学分析来检查工程化受体。结果将提供有关与 NMDA 受体激活相关的分子运动的基本信息，并将提供有关该受体药物作用的可能机制的宝贵见解。这些知识将构成合理控制 NMDA 受体活性所需的概念基础，作为中风、成瘾和精神疾病的治疗策略。公共健康相关性：该项目将提供有关 NMDA 受体变得活跃的机制的基本信息。目前缺乏这些信息，但合理设计急性和慢性神经退行性疾病（中风、阿尔茨海默病）、成瘾、慢性疼痛、认知障碍和精神疾病的新治疗方法需要这些信息。