Structure of the GABA-A Receptor Binding Sites

GABA-A 受体结合位点的结构

基本信息

批准号：
8640210
负责人：
CYNTHIA M CZAJKOWSKI
金额：
$ 49.84万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-12-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8640210
关键词：
Agonist Alzheimer&apos s Disease Anesthetics Anti-Anxiety Agents Antiemetics Barbiturates Benzodiazepines Binding Binding Sites Biochemical Brain Butyric Acids Chemicals Convulsants Coupling Crosslinker Cysteine Data Defect Disulfides Drug Design Drug effect disorder Drug usage Electron Spin Resonance Spectroscopy Emetics Ethanol Etiology GABA A Receptor Binding Gated Ion Channel Genes Glycine Receptors Goals HTR3A gene Homologous Gene Intravenous Ion Channel Ion Channel Gating Ion Channel Protein Knowledge Lead Length Ligand Binding Domain Ligands Lighting Maps Measures Mediating Membrane Mental disorders Metals Molecular Molecular Conformation Molecular Structure Motion Movement Muscle relaxants Neurons Neurotransmitters Nicotine Dependence Nicotinic Receptors Outcome Pectobacterium chrysanthemi Pharmaceutical Preparations Process Proteins Protons Radial Receptor Activation Relative (related person)Serotonin Signal Transduction Site Spin Labels Structural Models Structure Synapses Testing Therapeutic Thermodynamics Transmembrane Domain Vestibule Work barbituric acid salt crosslink design disease-causing mutation extracellular flexibility gamma-Aminobutyric Acid hypnotic improved insight member mutant nervous system disorder neurosteroids novel strategies prevent public health relevance receptor receptor binding research study sedative simulation voltage clamp

项目摘要

DESCRIPTION (provided by applicant): Chemical signaling in the brain relies on rapid opening and closing of ligand-gated ion channels (LGICs) in the membranes of nerve cells. Members of the pentameric LGIC superfamily include nicotinic acetylcholine receptors (nAChR), serotonin-type-3 receptors (5HT3R), gamma-amino butyric acid type A receptors (GABAAR) and glycine receptors. Defects in these channels lead to a variety of neurological diseases and psychiatric disorders and a number of therapeutic drugs, including muscle relaxants, sedative-hypnotics, anti- convulsants, anxiolytics, intravenous and volatile anesthetics, anti-emetics, drugs for nicotine addiction and drugs to treat Alzheimer's disease target these channels. For these receptors, binding of neurotransmitter in the extracellular ligand-binding domain triggers opening of an intrinsic ion channel more than 50¿ away in the transmembrane domain of the receptor. Although we know a fair amount about the structure of these receptors, the mechanisms by which the binding of neurotransmitter triggers channel opening are still under debate and our understanding of the protein motions underlying this process limited. The general plan of this proposal is to investigate the binding-to-gating motions in the prokaryotic pLGIC homologs from Gloeobacter violaceus (GLIC) using site-directed spin labels and electron paramagnetic resonance spectroscopy (SDSL- EPR) and to test these motions in the GABAAR using an array of biochemical and electrophysiological approaches including voltage clamping, mutant cycle analysis, cysteine cross-linking, disulfide trapping and structural modeling. We will focus on three key regions: the extracellular binding domain (EBD), the gating interface and the transmembrane channel domain (TCD). These studies will build on our previous work and will provide new insights into how neurotransmitters activate pLGICs and how allosteric drugs modulate their activity. A deeper understanding of how these channels work at a molecular level will improve our ability to predict the actions of drugs and ligands that act on these channels, design safer and more effective drugs, develop better therapeutic strategies, and understand the etiology of disease-causing mutations.

描述（由适用提供）：大脑中的化学信号传导依赖于神经细胞机理中配体门控离子通道（LGIC）的快速打开和关闭。五聚体LGIC超家族的成员包括烟碱乙酰胆碱受体（NACHR），5-羟色胺型-3受体（5HT3R），γ-氨基丁酸A型A型受体（GABAAR）和甘氨酸受体。 Defects in these channels lead to a variety of neurological diseases and psychiatric disorders and a number of therapeutic drugs, including muscle relaxants, sedative-hypnotics, anti-convulsants, anxiolytics, intravenous and volatile anesthetics, anti-emetys, drugs for nicotine addiction and drugs to treat Alzheimer's disease target these channels.对于这些受体，在细胞外配体结合结构域中神经递质的结合触发了固有离子通道的打开，而我们知道这些受体的结构相当多，而这些受体的结构相当多，神经递质触发通道的结合仍在蛋白质中，我们对蛋白质的理解仍然有限。该提案的一般计划是使用位于定位的自旋标签和电子顺磁共振光谱（SDSL- EPR）调查原核生物PLGIC同源物（Gloeobacter clic）（Glic）中的结合对基因运动，并在使用GABAAR中测试了这些动态，并使用包括生物化学的阵列进行了涉及这些运动的这些运动。半胱氨酸交联，二硫键捕获和结构建模。我们将重点关注三个关键区域：细胞外结合结构域（EBD），门控界面和跨膜通道域（TCD）。这些研究将以我们以前的工作为基础，并将提供有关神经递质如何激活PLGIC以及变构药物如何调节其活性的新见解。对这些通道如何在分子水平上起作用的更深入了解将提高我们预测在这些通道上作用，设计更安全和更有效的药物，发展更好的治疗策略并了解引起疾病突变的病因的药物和配体的作用的能力。