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: 10166355
• 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/10166355
• 关键词: Address; Biochemical; Biological; Biological Models; Biophysics; Complex; Cryoelectron Microscopy; Ion Channel; Methods; Molecular Conformation; Nociceptors; Pharmacology; Physiological; Protein Conformation; Protein Dynamics; Sampling; Signal Transduction; Stimulus; Structural Protein; Structure; TRP channel; TRPV1 gene; Technology; conformational conversion; macromolecule; novel; particle; 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）中的技术突破已启用原子 在前所未有的快速速度中，许多具有挑战性的生物大分子的结构确定。也是如此 促进了解决挑战性的结构生物学问题，否则难以解决。 除原子结构之外，功能状态之间的蛋白质动力学和构象过渡是 特别适合通过单个粒子冷冻EM研究的问题。 TRPV1离子通道是多型伤害感受器，它整合了从一系列生化和 生物物理刺激。它在生物化学上表现良好，在生理和药理学上的表征都很好。 具有各种内源性刺激和天然外源化合物，将通道捕获在各种功能中 国家，我们确定了各种稳定构象中TRPV1的结构。在下一步中，我们将使用 TRPV1作为模型系统，用于利用新型的冷冻EM进步来探测构象状态的范围 由这个复杂的信号积分器采样。对于许多其他TRP频道，没有合适的频道 药理学工具可将通道以特定的稳定构象捕获，并使用模型开发的方法 系统可用于研究通道门控机制。