Structural Studies of Ion Channel Assembly and Signaling Complexes

描述（由申请人提供）：该项目的长期目标是发展对离子通道功能和调控的高分辨率理解。我们专注于了解电压门控离子通道（VGIC）超家族的两种典型类别Kv7和TRPM通道的调制和组装的架构基础。这些通道的大分子复合物在整个神经、感觉、听觉和心血管系统的生物电信号传导中起着关键作用。努力的方向是了解来自这些通道的细胞内模块如何与调节蛋白相互作用，以及作为这两类的共同特征的卷曲线圈组装结构域如何直接组装和组装特异性决定因素。我们关注两个核心问题：1)Kv7 c端尾部钙调蛋白结合装置的结构性质是什么？疾病突变如何影响钙调蛋白与c端尾部的相互作用？2) TRPM和Kv7通道胞内组装域的结构性质是什么？是否存在指导异质复合物组装的共同主题？阐明潜在的Kv7和TRPM结构框架对于理解这些和其他vgic如何整合到细胞内信号通路以及开发干预控制通道功能的新方法至关重要。我们的工作包括多学科方法，包括生化，生物物理，x射线晶体学和电生理测量来解剖功能。由于其在人体生理中的重要作用，vgic成为治疗心律失常、高血压、充血性心力衰竭、癫痫和慢性疼痛的药物靶点。因此，在原子水平上详细了解它们的结构和作用机制，将极大地有助于开发针对各种人类疾病的有价值的治疗药物。

DESCRIPTION (provided by applicant): The long-term goals of this project are to develop a high-resolution understanding of ion channel function and regulation. We are focused on understanding the architectural foundations that underlie the modulation and assembly of two exemplar classes of the voltage-gated ion channel (VGIC) superfamily, Kv7 and TRPM channels. Macromolecular complexes of these channels play pivotal roles in bioelectrical signaling throughout the nervous, sensory, auditory, and cardiovascular systems. Efforts are directed at understanding how intracellular modules from these channels interact with regulatory proteins and how the coiled-coil assembly domains that are a common feature of both classes direct assembly and assembly specificity determinants. We focus on two central questions: 1) What is the structural nature of the calmodulin binding apparatus in the Kv7 C-terminal tail and how do disease mutations affect calmodulin interactions with the C-terminal tail? 2) What is the structural nature of the intracellular assembly domains of TRPM and Kv7 channels and are there common themes directing heteromeric complex assembly? Elaboration of the underlying Kv7 and TRPM structural framework is essential for understanding how these and other VGICs are integrated into intracellular signaling pathways and for developing novel ways to intervene to control channel function. Our efforts encompass a multidisciplinary approach that includes biochemical, biophysical, X-ray crystallographic, and electrophysiological measurements to dissect function. Because of their important role in human physiology, VGICs are the targets for drugs with great utility for the treatment of cardiac arrhythmias, hypertension, congestive heart failure, epilepsy, and chronic pain. Thus, understanding their structures and mechanisms of action at atomic level detail should greatly assist the development of valuable therapeutic agents for a wide range of human ailments.