Structural Studies of Voltage Gating in Voltage-Gated Sodium Channels

电压门控钠通道中电压门控的结构研究

基本信息

批准号：
7305564
负责人：
BENJAMIN W SPILLER
金额：
$ 29.17万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-15 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7305564
关键词：
Action Potentials Affect Anti-Arrhythmia Agents Antibodies Architecture Binding Biological Assay Calcium Calcium Channel Cells Chemicals Complex Crystallization Crystallography Data Data Set Dependence Dihydropyridines Disease Disulfides Electrons Engineering Epilepsy Escherichia coli Event Exhibits Fab Immunoglobulins Family Freezing Functional disorder Gated Ion Channel Health Benefit Homo Hypertension Ions L-Type Calcium Channels Label Lead Mammalian Cell Mechanics Medical Melanocytic nevus Membrane Methionine Methods Modeling Mole the mammal Molecular Molecular Conformation Monoclonal Antibodies Motion Mutate Mutation Permeability Pharmaceutical Preparations Phase Positioning Attribute Potassium Channel Property Public Health Purpose Research Personnel Resolution Role Sodium Sodium Channel Solutions Structure Therapeutic Tissues Work antibody engineering base chronic neuropathic pain dihydropyridine disease-causing mutation disulfide bond improved interest method development mutant physical model programs sensor voltage voltage gated channel

项目摘要

DESCRIPTION (provided by applicant): Action potentials are typically initiated by sodium channels (Navs). This role is shared by Navs in many tissues, and mutations or other dysfunction of Navs lead to tissue specific diseases, including arrythmias, epilepsy, and chronic neuropathic pain. A key similarity in all channelopathies is that an insult to the basic architecture of the channel leads to a change in function. Determining this basic architecture at atomic resolution and the stereo-chemical details of selectivity, gating, and of anti-hypertension drug binding are the purposes of this proposal. While structural studies of K+ channels are quite advanced, there are currently no analogous studies and no stereo-chemical understanding of Na+ and Ca2+ selectivity or of drug binding. The broad aim of this proposal is to determine the mechanisms of sodium and calcium selectivity and anti-hypertension drug binding in a family of voltage-gated ion channels. The first specific aim will complete our current structure, the second specific aim will determine the mechanism of ion selectivity for both Na+ and Ca2+, and the third aim will develop and utilize methods to stabilize inherently flexible molecules. Multiple antibodies and engineered disulfide bonds will be used to stabilize specific gating states. These states will be confirmed by structural studies, and their structures will be determined by x-ray crystallography. The long-term benefit of this work will be a unifying model for the hundreds of disease causing mutations, a stereo-chemical view of drug binding leading to improved therapeutics, a better understanding of ion selectivity and the relationship between Navs and Cavs, and a physical model for channel gating. An additional benefit will be the development of methods for immobilizing inherently flexible molecules to aid in crystallization. This project will use x-ray crystallography to determine the first Na+ channel structure. The public health benefit will be improved treatment methods for Na+ channel dysfunction resulting from improved anti- arrhythmias drugs.

描述（由申请人提供）：动作电位通常由钠通道（Navs）启动。许多组织中的 Navs 都具有这种作用，并且 Navs 的突变或其他功能障碍会导致组织特异性疾病，包括心律失常、癫痫和慢性神经性疼痛。所有通道病的一个关键相似之处在于，对通道基本结构的破坏会导致功能的改变。本提案的目的是确定原子分辨率的基本结构以及选择性、门控和抗高血压药物结合的立体化学细节。虽然 K+ 通道的结构研究相当先进，但目前还没有类似的研究，也没有对 Na+ 和 Ca2+ 选择性或药物结合的立体化学理解。该提案的主要目标是确定电压门控离子通道家族中钠和钙选择性以及抗高血压药物结合的机制。第一个具体目标将完成我们目前的结构，第二个具体目标将确定 Na+ 和 Ca2+ 的离子选择性机制，第三个目标将开发和利用稳定固有柔性分子的方法。多种抗体和工程二硫键将用于稳定特定的门控状态。这些状态将通过结构研究得到证实，并且它们的结构将通过X射线晶体学确定。这项工作的长期好处将是数百种引起疾病的突变的统一模型、药物结合的立体化学观点，从而改进治疗方法、更好地理解离子选择性以及 Navs 和 Cavs 之间的关系，以及通道门控的物理模型。另一个好处是开发固定固有柔性分子以帮助结晶的方法。该项目将使用 X 射线晶体学来确定第一个 Na+ 通道结构。改进的抗心律失常药物所导致的 Na+ 通道功能障碍的治疗方法将得到改善，这将给公众健康带来好处。