Investigation of the Role of the KCNH Voltage-Gated Potassium Channel Intracellular Domains in Gating

KCNH 电压门控钾通道细胞内结构域在门控中的作用研究

• 批准号: 10393985
• 负责人: Whitney Alexandra Stevens-Sostre
• 金额: $ 3.55万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10393985
• 关键词: Action Potentials; Address; Animal Model; Area; Arrhythmia; Award; Binding; Binding Sites; Biophysics; Brain; C-terminal; Calcium; Calmodulin; Cardiac; Cardiac Death; Collaborations; Complex; Couples; Critical Thinking; Cryoelectron Microscopy; Cyclic Nucleotides; Development; Disease; Doctor of Philosophy; Electrodes; Electron Microscope; Electrophysiology (science); Epilepsy; Ethers; Family; Functional disorder; Future; Future Generations; Gap Junctions; Genes; Goals; Heart; Human; Hydrophobic Interactions; Hydrophobicity; Image; Investigation; Ion Channel; Ion Channel Protein; Journals; Kinetics; Knowledge; Laboratories; Laboratory Research; Learning; Life; Light; Malignant Neoplasms; Maps; Measures; Mediating; Memory; Methods; Molecular; Molecular Machines; Mutation; N-terminal; Nature; Nervous system structure; Neurologic; Neurological Models; Neurons; Neurosciences; Nucleotides; Oral; Oxidation-Reduction; Oxygen; Pathologic Processes; Pathway Analysis; Pathway interactions; Peer Group; Pharmaceutical Preparations; Phase; Physiological; Physiological Processes; Point Mutation; Positioning Attribute; Potassium Channel; Presynaptic Terminals; Process; Property; Publications; Rattus; Research; Research Personnel; Research Project Grants; Role; Schizophrenia; Scientist; Shapes; Site-Directed Mutagenesis; Structure; Synaptic Transmission; Techniques; Testing; Thermodynamics; Training; Ventricular; Voltage-Gated Potassium Channel; Work; Xenopus oocyte; alpha helix; biophysical techniques; diagnostic biomarker; experience; insight; meetings; mutant; nervous system disorder; neuronal excitability; neurotransmitter release; novel; optogenetics; posters; protein protein interaction; sensor; simulation; skills; synaptic function; targeted treatment; therapeutic development; voltage; voltage clamp

PROJECT SUMMARY/ABSTRACT KCNH channels, also known as the Ether-à-go-go (Eag) family, are voltage-gated potassium ion channels that have roles in neuronal excitability and cardiac repolarization. The dysfunction of these ion channels is implicated in a variety of diseases, including cancer, epilepsy, and cardiac arrythmia, making them promising targets for both diagnostic markers and the development of therapeutic drugs. KCNH channels like EAG and human Ether-à-go-go–Related Gene (hERG) possess unique and highly conserved intracellular domains that have evolved to serve unique physiological roles. The N-terminal Per-Arnt-Sim (PAS) domain has diverse functions in nature: they serve as input modules that mediate protein-protein interactions and as redox potential, oxygen, and light sensors. The C-terminal cyclic nucleotide binding homology (CNBh) domain has structural homology to cyclic nucleotide binding (CNB) domains but lack the ability to bind nucleotides. The intracellular PAS and CNBh domains modulate gating in KCNH channels and are known from structural and functional studies to associate in a complex. Several disease-associated mutations are concentrated at the PAS-CNBhD interface, highlighting the physiological importance of their interaction. However, details of how these domains interact to allosterically regulate critical channel functions, such as slow deactivation in hERG and calmodulin (CaM) inhibition in EAG1, are not understood. This proposal seeks to investigate the role of the KCNH channel intracellular domains in gating by combining structural, computational, and electrophysiological approaches. In Aim 1 (PhD Progress), I describe the presence of a hydrophobic interaction among residues in the PAS-cap (residues1-25), the downstream globular PAS (residues 26-135), and the CNBh domain of a neighboring subunit that are critical for slow deactivation in hERG channels. In Aim 2 (F99 Phase), I propose to uncover the allosteric pathways that mediate Ca2+-CaM inhibition of EAG1 currents. Given the importance of KCNH channels in various physiological and pathological processes, we expect our novel findings to have broad implications in neuroscience and beyond. During the F99 Phase, I will continue my thesis work in my advisor's state-of-the art laboratory with the guidance and support of my colleagues and graduate trainee peer group. I will learn cutting-edge techniques and methods of analysis, as well as develop my critical thinking skills to interpret my findings. I will continue engaging in professional development activities, including journal clubs, seminars, and lab meetings. Finally, I will share my scientific findings through publications, and oral and poster presentations at scientific meetings. My long-term goal is to understand the effects of channelopathies within the nervous system. Thus, for the K00 phase, I have identified specific areas, concepts, and techniques I must develop to become a well-rounded researcher and expert in neurological channelopathies. I intend to utilize my expertise to establish a diverse and equitable academic research laboratory to train future generations of scientists.

项目摘要/摘要 KCNH通道，也称为Ether-à-go-go（EAG）家族，是电压门控钾离子通道 在神经元令人兴奋和心脏复极中起作用。这些离子通道的功能障碍是 在各种疾病中实施 诊断标记和热药的发展的靶标。像eag这样的KCNH频道 人类以太乙醚相关的基因（HERG）潜在的独特且高度构成的细胞内结构域 已经演变为发挥独特的生理作用。 N末端每端SIM（PAS）域具有潜水员 自然界的功能：它们是介导蛋白质 - 蛋白质相互作用的输入模块和氧化还原 电势，氧气和光传感器。 C末端循环核苷酸结合同源（CNBH）域具有 与环状核苷酸结合（CNB）结构域的结构同源性，但缺乏结合核苷酸的能力。这 细胞内PA和CNBH结构域在KCNH通道中调节门控，并从结构和 功能研究以在复合物中关联。几个与疾病相关的突变集中在 PAS-CNBHD接口，突出了它们相互作用的物理重要性。但是，详细信息如何 这些域与变构调节关键通道功能相互作用，例如HERG中的缓慢失活 EAG1中的钙调蛋白（CAM）抑制作用尚不清楚。该建议旨在调查 通过结合结构，计算和电生理学，KCNH通道在门控中的细胞内结构域 方法。在AIM 1（PHD进度）中，我描述了残留物之间存在疏水相互作用 PAS-CAP（残基1-25），下游全局PA（残基26-135）和A的CNBH结构域 邻近的亚基对于在HERG频道中缓慢停用至关重要。在AIM 2（F99阶段）中，我建议 发现介导Ca2+-CAM抑制EAG1电流的变构途径。考虑到重要性 在各种物理和病理过程中，KCNH渠道，我们希望我们的新发现有 神经科学及以后的广泛含义。 在F99阶段，我将继续我的论文工作 我的同事和研究生实习生小组的指导和支持。我将学习尖端技术 和分析方法，以及发展我的批判性思维能力来解释我的发现。我会继续 从事专业发展活动，包括期刊俱乐部，半人数和实验室会议。最后，我 将在科学会议上通过出版物以及口头和海报演讲分享我的科学发现。 我的长期目标是了解神经系统中通道病的影响。那是K00 阶段，我已经确定了我必须开发的特定领域，概念和技术，以成为一个全面的 神经通道病的研究人员兼专家。我打算利用我的专业知识来建立潜水员 和公平的学术研究实验室，以培训子孙后代的科学家。