Structural biology of the alpha4beta2 nicotinic acetylcholine receptor

α4β2烟碱乙酰胆碱受体的结构生物学

基本信息

批准号：
8681089
负责人：
Ryan E Hibbs
金额：
$ 20.14万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8681089
关键词：
Absence of pain sensation Agonist Alcohol consumption Attenuated Automobiles Behavior Binding Biochemical Brain Cause of Death Cell Line Cessation of life Chewing Gum Complex Crystallization Detergents Development Drug usage Electrophysiology (science)Exhibits Family Goals HIV Health Injury Investigation Ions Laboratories Ligands Lipid Bilayers Mediating Membrane Proteins Methods Molecular Molecular Conformation Murder Mus Neurodegenerative Disorders Neurotransmitter Receptor Nicotine Nicotine Dependence Nicotinic Receptors Persons Phase Play Positioning Attribute Postdoctoral Fellow Proteins Research Resolution Roentgen Rays Role Self Administration Site-Directed Mutagenesis Smoker Source Structure Suicide System Techniques Tobacco use United States Work X-Ray Crystallography base channel blockers desensitization design new therapeutic target nicotine replacement nicotinic receptor alpha4beta2 novel novel therapeutics overexpression receptor small molecule smoking cessation stoichiometry structural biology three dimensional structure

Absence of pain sensation Agonist Alcohol consumption Attenuated Automobiles Behavior Binding Biochemical Brain Cause of Death Cell Line Cessation of life Chewing Gum Complex Crystallization Detergents Development Drug usage Electrophysiology (science)Exhibits Family Goals HIV Health Injury Investigation Ions Laboratories Ligands Lipid Bilayers Mediating Membrane Proteins Methods Molecular Molecular Conformation Murder Mus Neurodegenerative Disorders Neurotransmitter Receptor Nicotine Nicotine Dependence Nicotinic Receptors Persons Phase Play Positioning Attribute Postdoctoral Fellow Proteins Research Resolution Roentgen Rays Role Self Administration Site-Directed Mutagenesis Smoker Source Structure Suicide System Techniques Tobacco use United States Work X-Ray Crystallography base channel blockers desensitization design new therapeutic target nicotine replacement nicotinic receptor alpha4beta2 novel novel therapeutics overexpression receptor small molecule smoking cessation stoichiometry structural biology three dimensional structure

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项目摘要

DESCRIPTION (provided by applicant): Nicotine addiction is the leading source of preventable death worldwide. The bottleneck in rational design of novel therapeutics for nicotine addiction is high-resolution structural information for nicotinic acetylcholine receptors. Specifically, the alpha4beta2 subtype of the nicotinic receptor plays an essential role in nicotine addiction. Nicotine selectively upregulates this neurotransmitter receptor in systems ranging from clonal cell lines to smokers' brains, and mice lacking the alpha4beta2 receptor exhibit dramatically attenuated nicotine self-administration behavior. The only specific therapy for smoking cessation distinct from nicotine replacement is a partial agonist for the alpha4beta2 nicotinic receptor. Unfortunately, these therapies are only modestly effective in helping people quit smoking. The goal of this proposal is to resolve this pressing biomedical problem by determining the structure of the alpha4beta2 nicotinic receptor by X-ray crystallography. The alpha4beta2 nicotinic receptor is a particularly daunting target for crystallographic studies because it is: (1) a eukaryotic membrane protein; (2) an obligate heteropentameric protein; and (3) its subunit stoichiometry is variable, being formed from either two alpha and three beta subunits, or three alpha and two beta subunits. This proposal is organized into two phases. In the first, exploratory (R21) phase, the goals are to (1) identify alpha4 and beta2 subunit constructs that combine to express at high levels and form stable pentameric receptors, (2) purify a single stoichiometric species of receptor and (3) obtain preliminary crystals. In the second (R33) phase, the goals are to (1) obtain well-diffracting crystals of the alpha4beta2 receptor and (2) determine its structure in complex with different classes of ligands.

描述（由申请人提供）：尼古丁成瘾是全球可预防死亡的主要来源。尼古丁成瘾的新型治疗剂合理设计的瓶颈是烟碱乙酰胆碱受体的高分辨率结构信息。具体而言，烟碱受体的α4Beta2亚型在尼古丁中起着至关重要的作用瘾。尼古丁在从克隆细胞系到吸烟者的大脑的系统中有选择地上调这种神经递质受体，缺乏α4Beta2受体的小鼠表现出巨大的尼古丁自我加入行为。与尼古丁替代不同的戒烟的唯一特定疗法是α4Beta2烟碱受体的部分激动剂。不幸的是，这些疗法仅在帮助人们戒烟方面有效。该建议的目的是通过X射线晶体学确定α4Beta2烟碱受体的结构来解决这种紧迫的生物医学问题。 Alpha4beta2烟碱受体是晶体学研究的特别令人生畏的靶标，因为它是：（1）真核膜蛋白；（2）强制性异脑蛋白；（3）其亚基化学计量学是可变的，由两个α和三个β亚基或三个α和两个β亚基形成。该提议分为两个阶段。在第一个探索性（R21）阶段中，目标是（1）识别α4和beta2亚基构建体，这些构建体结合以高水平表达并形成稳定的五聚体受体，（2）纯化受体的单个化学计量物种，（3）获得初步晶体。在第二个（R33）阶段，目标是（1）获得α4Beta2受体的良好分裂晶体，（2）确定其与不同类别的配体的复合物中的结构。