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: 10728394
• 负责人: Sharona E Gordon
• 金额: $ 11.35万
• 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/10728394
• 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.

摘要 离子通道是具有门控跨膜孔的完整膜蛋白， 离子沿其电化学梯度向下移动，以消除化学、机械和光学的影响， 信号转换成电信号。在这个提案中，我们利用一系列创新工具来破译 TRP离子通道的两个亚家族TRPV 1 -2和TRPM 2的变构门控机制。 TRP通道以其多模态门控而闻名，其中刺激模态如 热、冷、离子、脂质、营养物、其它蛋白质和各种天然产物（例如， 辣椒素、薄荷醇）进行别构整合以确定中心离子的活性， 导电孔每一种刺激形式调节感测的构象能量学， module.传感模块进而调节构象能量学和电导 的毛孔。感测模块可以耦合到孔、彼此耦合或两者。TRP 通道提供了一个理想的系统，在其中破译如何变构构象能量学 产生蛋白质功能，因为它们受到许多刺激方式的调节， 对它们的结构域和感知之间的对应关系的重要理解 模块。该提案的目标是首次测量构象 TRP通道传感结构域的能量学及其与孔和彼此的偶联， 解决TRP通道生物学中的紧迫问题。我们的长远目标是了解 离子通道中变构构象转变的基本主题。我们最近 技术进步结合荧光寿命成像显微镜（FLIM）和补丁， 钳位电生理学以测量孔中的构象能量学， 模块同时承诺朝着这一目标快速进展。