A Regulator for Eag Family Channels

描述(由申请人提供)：人类乙醚-a-go-go相关基因1(Hergl)通道功能障碍在心脏病中起着关键作用，心脏病是美国的主要死亡原因。Herg1在心脏中大量表达，其延迟整流钾电流有助于心脏动作电位的复极化。它是EAG(Ethera-a-Go-Go)通道家族中最具特色的成员。我们已经分析了Herg1和EAG家族的其他通道的基因序列，以确定两个可能的配体结合位点，从而将这些通道定性为孤儿受体。这项建议的总体目的是确定Hergl和EAG家族其他通道的细胞内调节因子，并表征这些调节因子如何调节通道门控。调节剂将通过高通量的细胞代谢物化学文库筛选和自内向外的膜片钳来识别。利用分子生物学、蛋白质生物化学和电生理学，我将确定通道调制如何影响门控特性，并确定调节器结合通道的位置。本方案中的实验将有助于我们理解这些通道在生理学和病理学中的作用。这些实验的另一个好处是发现了涉及中枢神经系统的途径，如学习和记忆，或疾病的基础过程，如心律失常和癌症。最终，这个受体位置可能成为一个潜在的药物靶点，以改善包括长QT综合征在内的许多疾病。这项研究的目标是确定一种被称为以太通道的蛋白家族的调节因子，这种通道在大脑和心脏中都有表达。编码这些通道的基因突变会导致心律失常和死亡。使用细胞内代谢物的文库筛选，我将识别这些通道的细胞内调节因子，并确定它们如何影响这些通道。

DESCRIPTION (provided by applicant): The human ether-a-go-go related gene 1 (Hergl) channel dysfunction plays a critical role in heart disease, a leading cause of death in the United States. Hergl is abundantly expressed in the heart, and its delayed rectifying potassium current contributes to the repolarization of the cardiac action potential. It is the bestcharacterized member of the ether-a-go-go (Eag) family of channels. We have analyzed the genetic sequence of Hergl, and other Eag family channels to identify two probable ligand-binding sites, and therefore characterize these channels as orphan receptors. The overall purpose of this proposal is to identify intracellular regulators for Hergl and other Eag family channels, and to characterize how these regulators modulate channel gating. Regulators will be identified with a high throughput chemical library screen of cellular metabolites, and inside-out patch clamp. Using molecular biology, protein biochemistry, and electrophysiology, I will determine how gating properties are affected by channel modulation and I will locate the site where the regulator binds the channel. The experiments in this proposal will contribute to our understanding of the role of these channels in physiology and pathology. Another benefit of these experiments is uncovering pathways involved in the central nervous system, such as learning and memory, or processes that underlie disease such as cardiac arrhythmia and cancer. Ultimately, this receptor site may be a potential drug target to ameliorate a number of diseases, including long QT syndrome. The goal of this study is to identify regulators for a family of proteins known as ether-a-go-go (Eag) channels, which are expressed throughout the brain and the heart. Mutations in the genes encoding these channels result in cardiac arrhythmias and death. Using a library screen of intracellular metabolites, I will identify intracellular regulators for these channels and determine how they affect these channels.