CLC-2 voltage-gated chloride channel structure and ligand recognition

CLC-2电压门控氯离子通道结构和配体识别

基本信息

批准号：
10391191
负责人：
Merritt C Maduke
金额：
$ 43.29万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-30 至 2024-03-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10391191
关键词：
Action Potentials Architecture Area Attention Biophysics Brain CLC Gene CLC-2 protein Calcium Cations Charge Chemicals Chloride Channels Chlorides Classification Complex Cryoelectron Microscopy Data Dependence Detection Electrophysiology (science)Family Fluorescence Goals Green Fluorescent Proteins Human body Image Investigation Ion Channel Ion Channel Gating Ions Kidney Labyrinth Ligands Membrane Proteins Molecular Molecular Conformation Molecular Sieve Chromatography Molecular Structure Movement Nervous System Physiology Neuraxis Neuroglia Neurons Play Positioning Attribute Potassium Potassium Channel Preparation Protocols documentation Reporting Research Research Project Grants Resolution Risk Role Sampling Shapes Signal Transduction Skeletal Muscle Sodium Channel Structure Variant base computer studies design fascinate inhibitor/antagonist insight nanomolar neurophysiology next generation programs reconstruction screening small molecule small molecule inhibitor tool voltage voltage gated channel

Action Potentials Architecture Area Attention Biophysics Brain CLC Gene CLC-2 protein Calcium Cations Charge Chemicals Chloride Channels Chlorides Classification Complex Cryoelectron Microscopy Data Dependence Detection Electrophysiology (science)Family Fluorescence Goals Green Fluorescent Proteins Human body Image Investigation Ion Channel Ion Channel Gating Ions Kidney Labyrinth Ligands Membrane Proteins Molecular Molecular Conformation Molecular Sieve Chromatography Molecular Structure Movement Nervous System Physiology Neuraxis Neuroglia Neurons Play Positioning Attribute Potassium Potassium Channel Preparation Protocols documentation Reporting Research Research Project Grants Resolution Risk Role Sampling Shapes Signal Transduction Skeletal Muscle Sodium Channel Structure Variant base computer studies design fascinate inhibitor/antagonist insight nanomolar neurophysiology next generation programs reconstruction screening small molecule small molecule inhibitor tool voltage voltage gated channel

展开

项目摘要

In the central nervous system (CNS), voltage-gated ion channels play central roles in shaping action-potential firing. The cation-selective voltage-gated channels – sodium (Na+), potassium (K+), and calcium (Ca2+)) channels – have received intense scrutiny over the past decades. In contrast, the chloride (Cl–)-selective voltage-gated channel, CLC-2, is less well understood, despite its broad expression in neurons and glia throughout the CNS. A complete understanding of CLC-2’s contribution to CNS function will include an understanding of CLC-2’s molecular structure. Structurally, CLC channels possess a unique double-barreled architecture and operate through distinct gating (opening/closing) mechanisms that differ markedly from those of the well-studied Na+, K+, and Ca2+ channels. In addition to providing a critical framework for studying CLC-2 channel gating and permeation mechanisms, another compelling rationale for determining the CLC-2 structure is its value for understanding ligand interactions and guiding design of small-molecule probes. Such probes would be of great value in investigations of CLC-2 neurophysiology. Accordingly, the goal of this R21 project is develop expression and purification protocols for CLC-2 and to use cryo-electron microscopy to determine CLC-2 structures in the absence and presence of the selective CLC-2 inhibitor AK-42.

在中枢神经系统（CNS）中，电压门控离子通道在塑造动作电位中起着核心作用射击。阳离子选择电压门控通道 - 钠（Na+），钾（K+）和钙（Ca2+））通道 - 在过去的几十年中，受到了严格的审查。相反，氯化物（CL - ） - 选择性电压门控 CLC-2频道的理解不太了解，使其在整个中枢神经系统中的神经元和神经胶质中的广泛表达。完全了解CLC-2对CNS功能的贡献将包括对Clc-2的了解分子结构。在结构上，CLC频道具有独特的双管架构和操作员通过独特的门控（开放/关闭）机制，这些机制与研究良好的Na+，K+，明显不同和CA2+通道。除了提供研究CLC-2通道门控和渗透机制，确定CLC-2结构的另一个引人注目的理由是其值了解配体相互作用并指导小分子问题的设计。这样的问题将很棒 CLC-2神经生理学的投资价值。根据这个R21项目的目标，正在发展表达和CLC-2的纯化方案并使用冷冻电子显微镜确定CLC-2结构选择性CLC-2抑制剂AK-42的不存在和存在。