Functional mechanisms and therapeutic potential of EAG channel regulators

Ether-a-go-go（EAG）钾离子通道是神经元电生理的重要调节因子， 兴奋性和癌症进展。EAG通道功能缺陷与 患有神经系统疾病和癌症尽管EAG的生理重要性 通道，EAG通道调节细胞内配体的分子机制， 临床相关的EAG通道调节剂是未知的。本提案的目的是 揭示细胞内配体调节EAG通道的分子机制 我们的实验室最近发现，并确定这些治疗潜力， 用于治疗与EAG通道相关的疾病的配体。具体目标1： 解决了细胞内Per-Arnt-Sim（PAS）和环核苷酸的X射线结构， 结合同源性（CNBH）结构域的EAG通道结合到最近确定的 配体，并进行配体与PAS结合的计算模拟， CNBH结构域，以揭示EAG通道调节的结构基础， 胞内配体。然后，结构性调查结果将被用作指导 EAG通道调节的分子机制的功能实验 配体。在具体目标2中，我们计划使用表面等离子体共振方法来识别 通过PAS和CNBH结构域影响通道功能的新型EAG通道配体 接口.然后，我们将使用电生理学来确定 PAS/CNBH结构域界面的增强或减弱， 调节EAG通道的功能。在Specific Aim 3中，我们计划使用组织 培养物和斑马鱼异种移植物模型，以测试所鉴定的 用于治疗癌症的调节剂。这些研究的结果将是至关重要的 了解EAG及相关ERG和ELK的基本调节机制 通道，并用于获得EAG通道调节剂的治疗潜力。

Ether-a-go-go (EAG) potassium selective channels are important regulators of neuronal excitability and cancer progression. Defects in EAG channel function are associated with neurological disorders and cancer. Despite the physiological importance of EAG channels, molecular mechanisms of EAG channel regulation by intracellular ligands and clinically relevant EAG channel regulators are not known. The goal of this proposal is to uncover molecular mechanisms of EAG channel regulation by intracellular ligands recently discovered by our laboratory and to determine a therapeutic potential of these ligands for treatment of diseases linked to EAG channels. In Specific Aim 1 we plan to solve X-ray structures of the intracellular Per-Arnt-Sim (PAS) and cyclic nucleotide- binding homology (CNBH) domains of EAG channels bound to the recently identified ligands and conduct computational simulations of the ligand binding to the PAS and CNBH domains to uncover the structural basis of EAG channel regulation by the intracellular ligands. The structural findings will be then used as a road map to guide functional experiments on the molecular mechanisms of EAG channel regulation by the ligands. In Specific Aim 2 we plan to use surface plasmon resonance method to identify novel EAG channel ligands that affect channel function through PAS and CNBH domain interface. We will then use electrophysiology to determine functional implications of strengthening or weakening of the PAS/CNBH domain interface by the identified regulators on the function of EAG channels. In Specific Aim 3 we plan to use tissue culture and zebrafish xenograft models to test therapeutic potential of the identified regulators for treatment of cancer. The results of these studies will be crucial for understanding fundamental regulatory mechanisms of EAG and related ERG and ELK channels, and for attaining therapeutic potential of EAG channel regulators.