3D Structure and Function of Neuronal Nicotinic Acetylcholine Receptors

神经元烟碱乙酰胆碱受体的 3D 结构和功能

• 批准号: 8538526
• 负责人: Ryan E Hibbs
• 金额: $ 24.03万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8538526
• 关键词: Alzheimer' s Disease; Basic Science; Binding; Biochemical; Biological Assay; Biophysical Process; Caenorhabditis elegans; Central Nervous System Diseases; Collaborations; Complement; Crystallization; Detergents; Drug Design; Electrophysiology (science); Family; Gated Ion Channel; Genes; Goals; Health; Homo; Human; Immunoglobulin Fragments; Intervention; Invertebrates; Ion Channel; Ions; Ligands; Link; Mediating; Medical; Mental Health; Mental disorders; Methods; Modeling; Modification; Molecular; Molecular Conformation; Nervous system structure; Neurodegenerative Disorders; Neurons; Nicotinic Receptors; Organism; Orthologous Gene; Parkinson Disease; Perfusion; Peripheral Nervous System; Phase; Physiological; Positioning Attribute; Postdoctoral Fellow; Preparation; Proteins; Public Health; Receptor Activation; Receptor Gene; Research; Research Project Grants; Resolution; Rest; Schizophrenia; Site-Directed Mutagenesis; Structure; Synaptic Transmission; Techniques; Testing; Therapeutic; Training; Transmembrane Domain; Work; addiction; base; blind; career; desensitization; design; experience; glutamate-gated chloride channel; innovation; insight; interest; member; neurotransmission; new therapeutic target; novel; patch clamp; receptor; screening; therapeutic target; three dimensional structure

Project Summary: Neuronal nicotinic acetylcholine receptors (nAChRs) are pentameric ligand-gated ion channels that mediate fast synaptic transmission and are important and novel therapeutic targets in neurodegenerative diseases and mental illness. Currently there is no atomic-resolution structure for any nicotinic receptor, and no receptor in this Cys-loop receptor superfamily has been structurally characterized in more than one conformation. The long-term objectives of the research are to better understand the conformational changes that underlie the transitions between different functional states and to develop reliable, atomic-resolution models of nAChRs relevant to structure-guided drug design. Toward these goals the following approach will be used. 1. Through receptor ortholog screening, identify an ¿7 receptor candidate for crystallization, and test conditions to optimize receptor stability in detergent. 2. Using fast-perfusion patch clamp and single channel analysis, test combinations of construct modifications and pharmacological probes to stabilize distinct receptor conformations. 3. Crystallize receptor in different conformational states and determine atomic-resolution structures of (a) a closed-resting receptor, (b) an open-activated receptor and (c) a closed-desensitized receptor. The structural studies will be complemented with functional assays that test new structure-based hypotheses regarding mechanisms of state transitions in the receptor. The results will have broad implications for the Cys-loop receptor family in general and ¿7 nAChRs specifically.

项目概述：神经元烟碱乙酰胆碱受体（nAChRs）是一种五聚体配体门控离子 通道，介导快速突触传递，是重要的和新的治疗靶点， 神经退行性疾病和精神疾病。目前还没有任何原子分辨率结构， 烟碱受体，并且在该Cys环受体超家族中没有受体的结构特征在于： 不止一种构象。研究的长期目标是更好地了解 不同功能状态之间转变的基础构象变化， nAChRs的原子分辨率模型与结构导向药物设计相关。为了实现这些目标， 将使用以下方法。1.通过受体直向同源物筛选，确定一个7受体候选者， 结晶和测试条件以优化洗涤剂中的受体稳定性。2.使用快速灌注贴片 钳和单通道分析，测试结构修改和药理学探针的组合， 稳定不同受体构象。3.使受体以不同的构象状态结晶， 确定（a）闭合静息受体，（B）开放激活受体和（c） 封闭脱敏受体结构研究将辅以功能测定， 基于结构的假设，关于受体中状态转换的机制。结果会有 对Cys环受体家族的广泛影响，特别是对7种nAChR的影响。