Structure and mechanism of extrasynaptic GABAA receptors

突触外GABAA受体的结构和机制

基本信息

批准号：
10431870
负责人：
Dagimhiwat Legesse
金额：
$ 3.53万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10431870
关键词：
Acute Affinity Agonist Alcohols Analgesics Anesthesia procedures Anesthetics Anions Anticonvulsants Anxiety Barbiturates Benzodiazepines Bicuculline Binding Binding Sites Biochemical Biological Biological Assay Biophysics Brain Cerebellum Chlorides Chronic Cognition Complex Cryoelectron Microscopy Cytoplasmic Granules Data Data Set Disease Drug Receptors Drug abuse Electrophysiology (science)Emotions Epilepsy Ethanol Exhibits Extracellular Space Foundations Future GTP-Binding Protein alpha Subunits, Gs Goals Grant Intoxication Ion Channel Gating Ions Knockout Mice Lead Ligands Lipids Liposomes Mediating Membrane Modeling Molecular Conformation Motor Muscimol Neuraxis Neurons Neurotransmitter Receptor Neurotransmitters Pharmaceutical Preparations Pharmacology Physiological Play Point Mutation Postpartum Depression Preparation Process Property Proteins Recombinants Research Resolution Risk Role Sampling Scaffolding Protein Seizures Sensory Signal Transduction Site Sleep Specificity Structure Synapses Synaptic Receptors Testing Wakefulness Work addiction antagonist biophysical properties experimental study gamma-Aminobutyric Acid interest lipid I nanodisk nervous system disorder neurosteroids neurotransmission particle patch clamp radioligand receptor receptor binding receptor structure function reconstitution response sedative therapeutic target

项目摘要

Project Summary: Type A γ-aminobutyric acid receptors (GABAA) are pentameric ligand-gated ion channels found abundantly in the central nervous system. The binding of GABA, the major brain inhibitory neurotransmitter, to GABAA receptors, accounts for ~30% of the fast synaptic signaling in the brain. These receptors are important for sensory and motor processing, central autonomic control, and cognition. While much of the research in GABAA receptor structure and function has focused on the subunit assemblies found at synapses, a tremendous diversity of pharmacologically important GABAA receptor subtypes are found outside of synapses. The goal of this proposal is to determine and validate high-resolution structures of extrasynaptic GABAA receptors bound to distinct drug classes of relevance to neurological disorders, anesthesia, addiction and drug abuse. These receptors are potentiated by mildly intoxicating concentrations of ethanol, have high affinity for neurosteroids and are targets of anesthetics, analgesics, and anticonvulsant drugs. In two Aims, I propose to optimize conditions for expression and purification of defined heteromeric GABAA receptor assemblies, refine sample preparation for cryo-electron microscopy, collect and process single particle cryo-EM datasets, and build and refine atomic models of receptor-drug complexes. High-resolution structural information for physiologically important but understudied subunit assemblies, in complex with allosteric and orthosteric-site ligands will broadly inform on principles of ligand recognition and subunit assembly. !

项目摘要： A型γ-氨基丁酸受体（GABAA）是发现的五聚合配体的离子通道几乎在中枢神经系统中。 GABA的结合，主要大脑抑制性神经递质与 GABAA受体，约占大脑快速突触信号的约30％。这些受体很重要用于感官和运动处理，中央自主控制和认知。而大部分研究 GABAA受体结构和功能集中在突触处的亚基组件上在突触之外发现了药物上重要的GABAA受体亚型的多样性。目标该建议是确定和验证与与与神经系统疾病，麻醉，成瘾和药物滥用的不同药物类别。这些受体通过轻度肠毒浓度的乙醇具有潜在的受体，对神经类固醇具有很高的亲和力并且是麻醉药，镇痛药和抗惊厥药的靶标。在两个目标中，我建议优化表达和纯化的条件和纯化定义的杂体GABAA受体组件，精炼样品准备冷冻电子显微镜，收集和处理单个粒子冷冻-EM数据集，并构建和受体毒物复合物的完善原子模型。高分辨率的结构信息重要但知识的亚基组件，与变构和正常位配体的复杂性将广泛告知配体认可和亚基大会的原则。呢