Advancing cryo-EM technology to address difficult biological questions

摘要 单粒子低温电子显微镜（cryo-EM）的技术突破使原子 许多具有挑战性的生物大分子的结构测定以前所未有的速度。它还 有助于解决具有挑战性的结构生物学问题，否则将难以解决。 除了原子结构，蛋白质动力学和功能状态之间的构象转变也是其中之一。 这些问题特别适合于通过单粒子cryo-EM来研究。 TRPV 1离子通道是多模式伤害感受器，其整合从一系列生化和免疫反应中产生的信号。 生物物理刺激它在生化方面表现良好，在生理上和生物学上都有很好的特征。 随着各种内源性刺激和天然外源性化合物捕获通道的各种功能， 状态下，我们已经确定了TRPV 1在各种稳定构象的结构。下一步，我们将使用 TRPV 1作为模型系统，利用新的cryo-EM进展探测构象状态的范围 由该复信号积分器采样。对于许多其他TRP通道，其中没有合适的 药理学工具，以特定的稳定构象捕获通道，与模型开发的方法 系统可用于研究通道门控机制。

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.