Conformational Energetics and Heterogeneity to Reveal Gating Mechanisms of TRPV and TRPM Ion Channels

构象能量学和异质性揭示 TRPV 和 TRPM 离子通道的门控机制

• 批准号: 10752849
• 负责人: Sharona E Gordon
• 金额: $ 19.96万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752849
• 关键词: Biology; Capsaicin; Cells; Chemicals; Coupled; Coupling; Electrophysiology (science); Environment; Goals; Heterogeneity; Integral Membrane Protein; Ion Channel; Ions; Lipids; Measures; Mechanics; Membrane; Menthol; Modality; Molecular; Molecular Conformation; Natural Products; Nutrient; Optics; Process; Proteins; Signal Transduction; Stimulus; System; TRP channel; TRPV1 gene; Time; conformational conversion; fluorescence lifetime imaging; innovation; multimodality; patch clamp; protein function; sensor; sulfated glycoprotein 2; tool

ABSTRACT Ion channels are integral membrane proteins with gated transmembrane pores that conduct ions down their electrochemical gradients to transduce chemical, mechanical, and optical signals into electrical signals. In this proposal, we leverage a host of innovative tools to decipher allosteric gating mechanisms in two subfamilies of TRP ion channels, TRPV1-2 and TRPM2. TRP channels are famous for their multimodal gating whereby stimulus modalities as diverse as heat, cold, ions, lipids, nutrients, other proteins, and a variety of natural products (e.g., capsaicin, menthol) are allosterically integrated to determine the activity of a central ion- conducting pore. Each stimulus modality regulates the conformational energetics of a sensing module. The sensing modules in turn regulate the conformational energetics and conductance of the pore. The sensing modules may be coupled to the pore, to each other, or both. TRP channels provide an ideal system in which to decipher how allosteric conformational energetics produce protein function because they are regulated by many stimulus modalities, and we have significant understanding of the correspondence between their structural domains and sensing modules. The goal of this proposal is to measure, for the first time, the conformational energetics of TRP channel sensing domains and their coupling to the pore and to each other to solve pressing questions in TRP channel biology. Our long-term vision is to understand the general themes that underlie allosteric conformational transitions in ion channels. Our recent technical advances combining fluorescence lifetime imaging microscopy (FLIM) and patch- clamp electrophysiology to measure conformational energetics in the pore and a sensing module simultaneously promise rapid progress toward this goal.

抽象的 离子通道是带有门控跨膜孔的整体膜蛋白 离子将其电化学梯度沿化学，机械和光学转化 信号进入电信号。在此提案中，我们利用许多创新工具来破译 TRPV1-2和TRPM2的两个亚家族中的变构门控机制。 TRP频道以其多模式门控而闻名，刺激方式多样化 热，冷，离子，脂质，营养，其他蛋白质和各种天然产物（例如， 辣椒素，薄荷酚是变构的，以确定中央离子的活性 进行孔。每种刺激方式调节传感的构象能量学 模块。传感模块反过来调节构象能量和电导 孔。传感模块可以耦合到孔，彼此或两者兼而有之。 trp 渠道提供了一个理想的系统，可以破译变构构象能力如何 产生蛋白质功能，因为它们受许多刺激方式调节，我们有 对它们的结构域和感应之间的对应关系的重大理解 模块。该提议的目的是首次衡量构象 TRP通道传感域的能量学及其耦合到孔，彼此之间 解决TRP渠道生物学中的紧迫问题。我们的长期愿景是了解 基于离子通道中变构构象转变的一般主题。我们最近 结合荧光寿命成像显微镜（FLIM）和斑块的技术进步 夹紧电生理学以测量孔中的构象能和感应 模块同时承诺朝着这个目标迅速进步。