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

• 批准号: 7683100
• 负责人: BENJAMIN W SPILLER
• 金额: $ 28.61万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7683100
• 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; 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; Research Personnel; Resolution; Role; Sodium; Sodium Channel; Solutions; Structure; Therapeutic; Tissues; Work; antibody engineering; base; chronic neuropathic pain; dihydropyridine; disease-causing mutation; disulfide bond; flexibility; improved; interest; method development; mutant; physical model; programs; sensor; voltage; voltage gated channel

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）：操作电位通常由钠通道（NAVS）发起。 NAV在许多组织中共有这种作用，而NAV的突变或其他功能障碍会导致组织特异性疾病，包括芳香族，癫痫和慢性神经性疼痛。所有通道病的关键相似性是对通道的基本体系结构的侮辱会导致功能的变化。在原子分辨率上确定这种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是该提议的目的。尽管K+通道的结构研究非常先进，但目前尚无类似研究，也没有对Na+和Ca2+选择性或药物结合的立体化学理解。该提案的广泛目的是确定在电压门控离子通道家族中钠和钙选择性和抗高血压药物结合的机制。第一个具体目标将完成我们当前的结构，第二个特定目标将确定Na+和Ca2+的离子选择性机制，而第三个目标将开发和利用方法来稳定固有的柔性分子。多种抗体和工程化的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确认，其结构将通过X射线晶体学确定。这项工作的长期益处将成为导致突变的数百种疾病的统一模型，这是对药物结合的立体化学观点，从而可以改善治疗疗法，对离子选择性以及NAVS和CAVS之间的关系有更好的了解以及通道门控的物理模型。另一个好处将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。公共卫生益处将改善因抗心律失常药物改善的Na+通道功能障碍的治疗方法。