Structural biology of neurotransmitter ion channels

神经递质离子通道的结构生物学

• 批准号: 8608553
• 负责人: James E Gouaux
• 金额: $ 27.72万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8608553
• 关键词: Achievement; Agonist; Alzheimer' s Disease; Anesthetics; Anticonvulsants; Basic Science; Binding; Binding Sites; Biological Assay; Biological Process; Butyric Acids; Caenorhabditis elegans; Cells; Chemicals; Communication; Complex; Crystallization; Detergents; Development; Disease; Environment; Epilepsy; Event; Family; Foundations; Gated Ion Channel; Glutamates; Glycine; Glycine Receptors; Heterogeneity; Human; Injury; Insecta; Integral Membrane Protein; Invertebrates; Ion Channel; Ion Channel Gating; Ion Channel Protein; Ligand Binding; Ligands; Light; Link; Lipids; Mammalian Cell; Maps; Measurement; Membrane; Membrane Proteins; Methods; Micelles; Molecular; Molecular Machines; Molecular Mechanisms of Action; Mutagenesis; Nervous system structure; Neurons; Neurotransmitters; Nicotine; P2X-receptor; Parkinson Disease; Peripheral Nervous System; Phase; Play; Porifera; Production; Publishing; Refractory; Resolution; Role; Serotonin; Signal Transduction; Site; Site-Directed Mutagenesis; Source; Structure; Testing; Therapeutic Agents; Work; design; drug discovery; extracellular; falls; glutamate-gated chloride channel; glycosylation; insight; novel; novel therapeutics; overexpression; receptor; receptor binding; screening; sedative; structural biology

DESCRIPTION (provided by applicant): The normal development and function of the human nervous system is critically dependent upon the function of neurotransmitter- or ligand-gated ion channels, molecular machines which facilitate the communication between one nerve cell and another. Equally important, these ion channels are the targets of many therapeutic agents, from sedatives and anesthetics to anticonvulsants, and they are implicated or associated with a broad range of devastating diseases including Alzheimer's and Parkinson's diseases as well as epilepsy. Two major and important families of neurotransmitter-gated ion channels are the ATP- sensitive P2X receptors and the pentameric Cys-loop receptors, the latter of which subsumes a super family of receptors that includes those for 3-amino butyric acid (GABA), glycine, serotonin and, in invertebrates, for glutamate. Unfortunately there are no high resolution, atomic views of any of these receptors in complexes with their cognate neurotransmitters or agonists. Furthermore, in the case of Cys- loop receptors, there is not yet even a high resolution structure of a eukaryotic receptor or a published method for production of receptor suitable for high resolution structural studies. Without this information, we are unable to understand how these important neurotransmitter-gated ion channels 'work' and, most significantly, we do not have molecular maps of the binding sites for neurotransmitters on the one hand, and molecules that inhibit neurotransmitter activity, i.e. antagonists, on the other hand. Because neurotransmitter-gated ion channels are multimeric integral membrane proteins they are particularly difficult to isolate and crystallize. The aim of the work proposed in this application is to develop new methods to screen and prepare neurotransmitter-gated ion channels for crystallographic studies, to design and implement new crystallization screening conditions, and to apply these methods to study P2X and Cys-loop receptors. Most specifically, we aim to solve high resolution x-ray crystal structures of P2X and Cys-loop receptors bound to their cognate neurotransmitter and to competitive antagonists, to test the veracity of the mapped sites by site-directed mutagenesis and ligand-binding assays, and to develop molecular mechanisms for the action of agonists and antagonists in these receptors. Taken together, our studies will provide the first atomic resolution views of these crucial receptors in their active and inhibited states, thus not only providing fundamental insight into their biological function, but also laying the foundation for the rational design of new therapeutic agents.

描述（由申请人提供）：人类神经系统的正常发育和功能在严重取决于神经递质或配体门控离子通道的功能，即促进一个神经细胞和另一个神经细胞之间通信的分子机器。同样重要的是，这些离子通道是许多治疗剂的靶标，从镇静剂和麻醉剂到抗惊厥药，它们与包括阿尔茨海默氏病和帕金森氏病以及癫痫病的广泛毁灭性疾病有关或与广泛的毁灭性疾病有关。神经递质门控离子通道的两个主要且重要的家族是ATP敏感的P2X受体和五聚体Cys-Loop受体，后者涵盖了包括3-氨基丁酸（GABA），甘氨酸），甲状腺素，音出舌蛋白和内覆盖物的超级家族的受体家族。不幸的是，在复合物中，没有高分辨率，原子观点与它们的同源神经递质或激动剂。此外，在Cys-Loop受体的情况下，甚至还没有真核受体的高分辨率结构或用于生产适合高分辨率结构研究受体的已发表方法。没有这些信息，我们将无法理解这些重要的神经递质门控离子通道的“工作”，最重要的是，我们一方面没有神经递质的结合位点的分子图，而分子抑制神经递质活性，即拮抗剂，即其他手动。由于神经递质门控离子通道是多聚体整合膜蛋白，因此特别难以隔离和结晶。本应用程序提出的工作的目的是开发新方法来筛选和准备神经递质门控的离子通道进行晶体学研究，设计和实施新的结晶筛查条件，并应用这些方法研究P2X和CYS-loop受体。最具体地说，我们旨在解决与它们的同源神经递质和竞争性拮抗剂结合的p2x和Cys-loop受体的高分辨率X射线晶体结构，以通过位置为导向的诱变和配体结合分析测试映射位点的真实性，并开发出分子机制的Agonists和Agonistrists和这些受体机构。综上所述，我们的研究将在其活跃和抑制的状态中提供这些关键受体的第一个原子分辨率观点，因此不仅提供了对其生物学功能的基本见解，而且还为新治疗剂的合理设计奠定了基础。