STRUCTURE AND MECHANISM OF A POLYMODAL TRP ION CHANNEL

多峰TRP离子通道的结构和机制

基本信息

批准号：
9381325
负责人：
Peng Yuan
金额：
$ 33.36万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-15 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9381325
关键词：
Address Agonist Area Biochemical Biological Assay Biological Process Biology Biophysics Blood Vessels Calcium Calcium Signaling Chemicals Clinical Complement Complex Cryoelectron Microscopy Crystallization Data Detergents Development Disease Drug Targeting Drug effect disorder Electrophysiology (science)Foundations Functional disorder Goals Homologous Gene Hypertension Inflammation Inherited Integral Membrane Protein Intervention Ion Channel Ions Length Ligands Malignant Neoplasms Membrane Membrane Lipids Membrane Proteins Methods Molecular Mutation Nerve Degeneration Neurobiology Nociception Non-Insulin-Dependent Diabetes Mellitus Obesity Osmoregulation Pain Pathologic Permeability Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Physiological Physiological Processes Physiology Play Positioning Attribute Property Regulation Research Resolution Roentgen Rays Role Sensory Signal Transduction Site-Directed Mutagenesis Stimulus Structural Protein Structure Temperature Therapeutic Intervention Thermogenesis Work X ray diffraction analysis X-Ray Crystallography X-Ray Diffraction chronic pain design drug discovery high dimensionality human disease insight mechanical force member nervous system disorder novel novel therapeutics particle patch clamp receptor screening skeletal dysplasia structural biology three dimensional structure voltage

项目摘要

Structure and Mechanism of a Polymodal TRP Ion Channel Transient receptor potential (TRP) ion channels are crucial for sensory transduction and cellular signaling, and TRP channel dysfunction is associated with a vast array of hereditary and acquired diseases including cancer, chronic pain, hypertension, and neurological disorders. Because of their central roles in physiology and pathophysiology, TRP channels have been intensively studied and are among the most aggressively pursued drug targets. However, advances in our understanding of TRP channel function and therapeutic interventions have been hindered by a lack of three-dimensional high-resolution structural information for most TRP channels. Our long-term goal is to develop structural and biochemical approaches to elucidate molecular mechanisms of TRP channels at the atomic level. By developing new methods to systematically evaluate heterologous expression, purification, and optimization of TRP channel homologs, we have recently crystallized a nearly full-length functional channel and obtained preliminary X-ray diffraction to 4.8 Å resolution. With this technical breakthrough and further optimization, we are now able to combine X-ray crystallography, single-particle cryo-electron microscopy (cryo-EM), patch-clamp electrophysiology, and site-directed mutagenesis to address fundamental molecular mechanisms. Specifically, we aim to determine high-resolution X-ray and cryo-EM structures of the channel bound with agonists or antagonists, and in complex with membrane lipids that regulate channel activity, and to dissect the underlying mechanisms of disease-associated mutations. Our proposed work will provide X-ray and cryo-EM structures of a TRP channel in multiple functional states and uncover structural and molecular mechanisms. In doing so, we will not only bring fundamental insights into TRP channel function, but also establish a foundation for rational design of new therapeutics for the treatment of many channel-associated diseases.

多聚trp离子通道瞬态接收器电位（TRP）离子通道的结构和机制对于感觉转导和细胞信号传导至关重要，而TRP通道功能障碍与众多遗传性疾病和获得性疾病有关，包括癌症，慢性疼痛，高血压，高血压，高血压和神经系统疾病。由于它们在生理学和病理生理学中的主要作用，TRP通道一直是研究的，并且是最积极的药物靶标之一。但是，我们对TRP渠道功能和治疗干预措施的理解的进步对于大多数TRP通道缺乏三维高分辨率结构信息，这受到了阻碍。我们的长期目标是开发结构和生化方法，以阐明原子水平的TRP通道的分子机制。通过开发新方法来系统地评估TRP通道同源物的异源表达，纯化和优化，我们最近结晶了几乎全长的功能通道，并获得了初步的X射线衍射至4.8Å分辨率。通过这种技术突破和进一步的优化，我们现在能够将X射线晶体学，单粒子冷冻电子显微镜（Cryo-EM），斑块钳电生理学和定向诱变结合在一起，以解决基本分子机制。具体而言，我们旨在确定与激动剂或拮抗剂结合的通道的高分辨率X射线和冷冻EM结构，并与调节通道活性并剖析疾病相关突变的潜在机制的膜脂质。我们提出的工作将在多个功能状态以及揭示结构和分子机制中提供TRP通道的X射线和冷冻EM结构。这样一来，我们不仅会将基本见解带入TRP渠道功能，还为合理设计的新疗法建立了基础，以治疗许多渠道相关疾病。