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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：
9010982
负责人：
CYNTHIA M CZAJKOWSKI
金额：
$ 50.34万
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/9010982
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 Electrophysiology (science)Emetics Ethanol Etiology GABA A Receptor Binding Gated Ion Channel Genes Glycine Receptors Goals HTR3A gene Health 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 Serotonin Signal Transduction Site Spin Labels Structural Models Structure Synapses Testing Therapeutic Thermodynamics Transmembrane Domain Vestibule Work crosslink design disease-causing mutation extracellular flexibility gamma-Aminobutyric Acid hypnotic improved insight member mutant nervous system disorder neurosteroids novel strategies prevent 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（由申请人提供）：大脑中的化学信号依赖于神经细胞膜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）、血清素-3型受体（5HT3R）、γ -氨基丁酸A型受体（GABAAR）和甘氨酸受体。这些通道的缺陷导致各种神经系统疾病和精神疾病，许多治疗药物，包括肌肉松弛剂、镇静催眠药、抗惊厥药、抗焦虑药、静脉和挥发性麻醉剂、止吐药、尼古丁成瘾药物和治疗阿尔茨海默病的药物都针对这些通道。对于这些受体，神经递质在细胞外配体结合区域的结合触发了受体跨膜区域内超过50°的内在离子通道的打开。尽管我们对这些受体的结构有相当多的了解，但神经递质结合触发通道打开的机制仍在争论中，我们对这一过程背后的蛋白质运动的理解有限。本提案的总体计划是利用定向自旋标记和电子顺磁共振波谱（SDSL- EPR）研究紫Gloeobacter violaceus (GLIC) pLGIC同源原核生物的结合-门控运动，并利用一系列生化和电生理学方法，包括电压夹紧、突变周期分析、半胱氨酸交联、二硫捕获和结构建模，在GABAAR中测试这些运动。我们将重点关注三个关键区域：细胞外结合域（EBD），门控界面和跨膜通道域（TCD）。这些研究将建立在我们之前的工作基础上，并将为神经递质如何激活plgic以及变构药物如何调节其活性提供新的见解。更深入地了解这些通道在分子水平上是如何工作的，将提高我们预测作用于这些通道的药物和配体的作用，设计更安全、更有效的药物，制定更好的治疗策略，并了解致病突变的病因学的能力。

项目成果

期刊论文数量（35）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The GABAA receptor alpha 1 subunit Pro174-Asp191 segment is involved in GABA binding and channel gating.

GABAA 受体 α1 亚基 Pro174-Asp191 片段参与 GABA 结合和通道门控。

DOI：
10.1074/jbc.m211905200
发表时间：
2003
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Newell,JGlen;Czajkowski,Cynthia
通讯作者：
Czajkowski,Cynthia

Stoichiometry of expressed alpha(4)beta(2)delta gamma-aminobutyric acid type A receptors depends on the ratio of subunit cDNA transfected.

表达的α(4)β(2)δγ-氨基丁酸A型受体的化学计量取决于转染的亚基cDNA的比例。

DOI：
10.1074/jbc.m110.104257
发表时间：
2010
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Wagoner,KellyR;Czajkowski,Cynthia
通讯作者：
Czajkowski,Cynthia

Site-directed spin labeling reveals pentameric ligand-gated ion channel gating motions.

DOI：
10.1371/journal.pbio.1001714
发表时间：
2013-11
期刊：
PLoS biology
影响因子：
9.8
作者：
Dellisanti CD;Ghosh B;Hanson SM;Raspanti JM;Grant VA;Diarra GM;Schuh AM;Satyshur K;Klug CS;Czajkowski C
通讯作者：
Czajkowski C

Structural determinants for high-affinity zolpidem binding to GABA-A receptors.

高亲和力唑吡坦与 GABA-A 受体结合的结构决定因素。

DOI：
10.1124/mol.106.029595
发表时间：
2007
期刊：
Molecular pharmacology
影响因子：
3.6
作者：
Sancar,Feyza;Ericksen,SpencerS;Kucken,AmyM;Teissére,JeremyA;Czajkowski,Cynthia
通讯作者：
Czajkowski,Cynthia

Local and global ligand-induced changes in the structure of the GABA(A) receptor.

局部和整体配体诱导的 GABA(A) 受体结构变化。