Functional Architecture of Glutamate Receptor Channels

谷氨酸受体通道的功能结构

• 批准号: 6765167
• 负责人: LONNIE P WOLLMUTH
• 金额: $ 25.59万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6765167
• 关键词: AMPA receptors; NMDA receptors; Xenopus oocyte; biological signal transduction; brain; cell line; glutamate receptor; inhibitor /antagonist; ion channel blocker; membrane channels; neurophysiology; polymerase chain reaction; protein structure function; single cell analysis; site directed mutagenesis; synapses; voltage /patch clamp

DESCRIPTION (provided by applicant): The long-term objective of my laboratory is to understand the molecular mechanisms underlying signaling in the brain, especially as it relates to higher brain functions and disease states. My laboratory focuses mainly on synaptic physiology, specifically those synapses that use glutamate as their neurotransmitter. Glutamate receptors (GluR) mediate basic information processing in the brain and contribute to the cellular and molecular mechanisms underlying learning and memory, the development and maintenance of cellular connections, and pain transduction and perception. They have also been implicated in acute and chronic celt death, including that associated with numerous neurological diseases. The goal of the present proposal is to investigate the structural basis of channel gating in GluRs. Such information is key to understanding the fundamental role of GluRs in brain function as well as the development of drugs that attenuate the cell death they mediate under pathological conditions. This issue will be studied in recombinant GluRs using a variety of techniques including site-directed mutagenesis, cysteine substitutions, channel blockers, fast agonist application, and whole cell and single channel analysis. Aim 1 will study the structure of the extracellular vestibule in GluR channels, including how it relates to an asymmetry between subunits. Given the functional significance of the extracellular vestibule--it contains sites for pore blockers with therapeutic potential--defining this domain is essential. This work also forms the basis for Aim2, where we will study more specifically the conformational changes in the extracellular vestibule during gating. These conformational changes couple ligand binding to channel opening and therefore represent fundamental features of their function. Aim3 will further pursue issue related to channel gating specifically to desensitization. These experiments will provide key insights into an important physiological process. This work will define fundamental principles of structure/function in GluR channels and provide new tools and insights into means to attenuate the cell death GluR mediated under pathological conditions.

描述（由申请人提供）：我实验室的长期目标是了解大脑信号传导的分子机制，特别是与高级大脑功能和疾病状态相关的分子机制。我的实验室主要关注突触生理学，特别是那些使用谷氨酸作为神经递质的突触。谷氨酸受体 (GluR) 介导大脑中的基本信息处理，并有助于学习和记忆、细胞连接的发展和维持以及疼痛传导和感知的细胞和分子机制。它们还与急性和慢性细胞死亡有关，包括与许多神经系统疾病相关的死亡。本提案的目标是研究 GluR 通道门控的结构基础。这些信息对于理解 GluR 在大脑功能中的基本作用以及开发减轻其在病理条件下介导的细胞死亡的药物至关重要。将使用多种技术在重组 GluR 中研究这个问题，包括定点诱变、半胱氨酸取代、通道阻断剂、快速激动剂应用以及全细胞和单通道分析。目标 1 将研究 GluR 通道中细胞外前庭的结构，包括它与亚基之间的不对称性的关系。鉴于细胞外前庭的功能重要性——它包含具有治疗潜力的孔阻滞剂位点——定义这个结构域是至关重要的。这项工作也构成了 Aim2 的基础，我们将更具体地研究门控过程中细胞外前庭的构象变化。这些构象变化将配体结合到通道开放上，因此代表了其功能的基本特征。 Aim3 将进一步解决与通道门控相关的问题，特别是脱敏。这些实验将为重要的生理过程提供重要的见解。这项工作将定义 GluR 通道结构/功能的基本原理，并为减轻病理条件下 GluR 介导的细胞死亡的方法提供新的工具和见解。