STRUCTURE, FUNCTION, AND MODULATION OF SERORTONIN (3A) RECEPTORS

5-羟色胺 (3A) 受体的结构、功能和调节

• 批准号: 9898063
• 负责人: Sudha Chakrapani
• 金额: $ 7.16万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9898063
• 关键词: Biochemical; Bipolar Depression; Bipolar Disorder; Cancer Etiology; Clinical; Complement; Cryoelectron Microscopy; Development; Diarrhea; Drug Design; Drug Targeting; Electrophysiology (science); Environment; Event; Foundations; Gated Ion Channel; Goals; Infection; Ion Channel Gating; Irritable Bowel Syndrome; Length; Ligands; Lipids; Membrane; Membrane Lipids; Molecular; Molecular Conformation; Mood Disorders; Nausea and Vomiting; Neurologic; Pain; Physiologic pulse; Physiological; Play; Process; Proteins; Resolution; Rest; Role; Serotonin; Signal Transduction; Structure; Therapeutic; Treatment Efficacy; Work; cancer therapy; gastrointestinal; gastrointestinal function; inhibitor/antagonist; interdisciplinary approach; pain perception; particle; receptor; serotonin receptor; transmission process

Project Summary/Abstract The overarching goal of this proposal is to determine the structural basis for allosteric mechanisms governing gating and modulation in serotonin receptors (5-HT3AR). 5-HT3ARs play a crucial role in gastrointestinal functions, pain transmission, and mood disorders. Inhibitors of 5-HT3AR are in the clinical use to alleviate nausea and vomiting caused by cancer therapies, manage post-infection diarrhea and irritable bowel syndrome, and treat neurological conditions such as bipolar disorder and depression. Rational drug design has been limited by poor understanding of the structure-function correlations in 5-HT3AR and the downstream signaling events. We recently solved the structure of the full-length 5-HT3AR in the resting conformation by single-particle cryo- electron microscopy (cryo-EM). By capitalizing on this technical advancement and further biochemical optimization, we aim to determine the conformational changes underlying 5-HT3AR gating and lipid modulation. We will achieve these goals by a combination of multidisciplinary approaches that include cryo-EM, pulsed- EPR, and electrophysiology. Specifically, we will determine high-resolution snapshots of 5-HT3AR in multiple functional states, in modulator-bound conformations, and in the presence of membrane lipid constituents. These structures will be complemented with protein dynamic studies in a membrane environment and extensive functional analysis. Our proposed work is expected to provide structural blueprints of the channel in physiologically relevant conformations and unravel the molecular mechanisms underlying channel function. These studies will be foundational in the targeted drug design for safer and effective therapeutics.

项目摘要/摘要 该提案的总体目标是确定构构机制的结构基础 5-羟色胺受体（5-HT3AR）的门控和调节。 5-HT3AR在胃肠道中起着至关重要的作用 功能，疼痛传播和情绪障碍。 5-HT3AR的抑制剂可缓解恶心 以及由癌症疗法引起的呕吐，管理感染后的腹泻和肠易激综合症，以及 治疗神经系统疾病，例如躁郁症和抑郁症。理性药物设计受到限制 对5-HT3AR和下游信号事件的结构功能相关性的了解不足。我们 最近解决了全长5-HT3AR的结构，在单粒子冷冻的静止构象中 电子显微镜（冷冻EM）。通过利用这一技术进步并进一步的生化 优化，我们旨在确定5-HT3AR门控和脂质调制的构象变化。 我们将通过包括冷冻EM，脉冲 - EPR和电生理学。具体而言，我们将在多个中确定5-HT3AR的高分辨率快照 功能状态，在调节剂结合的构象中，并在膜脂质成分的存在下。这些 结构将在膜环境中与蛋白质动态研究相辅相成，广泛 功能分析。我们提出的工作有望在渠道中提供结构性蓝图 生理上相关的构象并揭示了基础通道功能的分子机制。 这些研究将是针对性药物设计的基础，以进行更安全，有效的治疗剂。