Structural Studies of Ion Channel Assembly and Signaling Complexes

描述（由申请人提供）：本项目的长期目标是发展对离子通道功能和调节的高分辨率理解。我们专注于理解的架构基础的调制和组装的两个范例类的电压门控离子通道（VGIC）超家族，Kv 7和TRPM通道。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中发挥关键作用。努力是针对了解如何从这些通道的细胞内模块与调控蛋白相互作用，以及如何卷曲螺旋组装域，这是一个共同的特点，这两个类直接组装和组装特异性决定因素。我们关注两个中心问题：1）Kv 7 C-末端尾中钙调素结合装置的结构性质是什么？疾病突变如何影响钙调素与C-末端尾的相互作用？2)TRPM和Kv 7通道的胞内组装结构域的结构性质是什么？是否存在指导异聚体复合物组装的共同主题？潜在的Kv 7和TRPM结构框架的识别对于理解这些和其他VGIC如何整合到细胞内信号传导通路中以及开发干预控制通道功能的新方法至关重要。我们的努力包括一个多学科的方法，包括生物化学，生物物理，X射线晶体学，和电生理测量解剖功能。由于其在人体生理学中的重要作用，VGIC是用于治疗心律失常、高血压、充血性心力衰竭、癫痫和慢性疼痛的药物的靶标。因此，在原子水平上详细了解它们的结构和作用机制，将大大有助于开发有价值的治疗药物，用于治疗各种人类疾病。

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.