Advancing cryo-EM technology to address difficult biological questions

推进冷冻电镜技术解决棘手的生物学问题

• 批准号: 10570241
• 负责人: Yifan Cheng
• 金额: $ 32.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10570241
• 关键词: Address; Biochemical; Biological; Biological Models; Biophysics; Complex; Cryoelectron Microscopy; Ion Channel; Methods; Molecular Conformation; Nociceptors; Physiological; Protein Conformation; Protein Dynamics; Sampling; Signal Transduction; Stimulus; Structure; TRP channel; TRPV1 gene; Technology; conformational conversion; macromolecule; novel; particle; pharmacologic; structural biology; tool

ABSTRACT Technological breakthroughs in single particle cryo-electron microscopy (cryo-EM) have enabled atomic structure determinations of many challenging biological macromolecules in an unprecedented rapid pace. It also facilitated addressing challenging structural biology questions that would otherwise be difficult to address. Beyond atomic structures, protein dynamics and conformational transitions between functional states are among questions that are particularly suited to be studied by single particle cryo-EM. TRPV1 ion channel is polymodal nociceptor that integrates signals generated from a range of biochemical and biophysical stimuli. It is biochemically well-behaved and physiologically and pharmacologically well characterized. With various endogenous stimuli and natural exogenous compounds trapping the channel in various functional states, we have determined structures of TRPV1 in various stable conformations. In the next step, we will use TRPV1 as a model system to exploit using novel cryo-EM advances to probe the range of conformational states sampled by this complex signal integrator. For many other TRP channels, in which there is no suitable pharmacological tools to trap the channels in specific stable conformations, methods developed with the model system can be used to study mechanisms of channel gating.

抽象的 单粒子冷冻电子显微镜 (cryo-EM) 的技术突破使原子 以前所未有的速度测定许多具有挑战性的生物大分子的结构。它还 促进解决具有挑战性的结构生物学问题，否则这些问题将难以解决。 除了原子结构之外，蛋白质动力学和功能状态之间的构象转变也是其中之一 特别适合通过单粒子冷冻电镜研究的问题。 TRPV1 离子通道是多模式伤害感受器，集成了一系列生化和 生物物理刺激。它的生化性能良好，生理学和药理学特征良好。 各种内源性刺激和天然外源性化合物在各种功能中捕获通道 状态，我们已经确定了 TRPV1 各种稳定构象的结构。在下一步中，我们将使用 TRPV1 作为模型系统，利用新颖的冷冻电镜技术进步来探测构象状态的范围 由该复杂信号积分器采样。对于许多其他 TRP 通道，其中没有合适的 将通道捕获到特定稳定构象的药理学工具，使用该模型开发的方法 系统可用于研究通道门控机制。