Towards a structural and temporal understanding of phototransduction

对光转导的结构和时间理解

基本信息

批准号：
7693695
负责人：
Paul S Park
金额：
$ 24.9万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7693695
关键词：
Abbreviations Address Adopted Architecture Area Artificial Membranes Atomic Force Microscopy Binding Biochemical Biochemical Genetics Biochemical Pathway Biogenesis Biological Biological Assay Biological Models Biology Bioluminescence Blindness Cell Line Cell Surface Proteins Cells Chimeric Proteins Complex Cyan Fluorescent Protein DNA Sequence Rearrangement Data Detection Development Diabetes Mellitus Dimensions Disease Dithiothreitol Drug Delivery Systems Electron Microscopy Electrophysiology (science)Energy Transfer Ethylmaleimide Event Exhibits Fluorescence Microscopy Fluorescence Resonance Energy Transfer Foundations Functional disorder G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors Genes Genetic Green Fluorescent Proteins Heart Diseases Housing Image Individual Information Systems Investigation Ions Knowledge Lead Life Light Lipids Location Maintenance Marketing Measures Mediating Membrane Membrane Proteins Mentors Methodology Methods Modification Molecular Molecular Biology Monitor Movement Nature Noise Optical Methods Organelles Pathway interactions Phase Phototransduction Physiological Processes Play Process Property Protein Family Proteins Research Resolution Retinal Dystrophy Rhodopsin Rod Outer Segments Role Scheme Signal Pathway Signal Transduction Signaling Molecule Signaling Protein Solutions Spectrum Analysis Staging Structure System Temperature Testing Therapeutic Time Transgenic Organisms Update Variant Vision Visual system structure Work Xenopus laevis addiction cell fixing computerized data processing dimer electron tomography fluorescence imaging in vivo innovation insight interest lecithin-retinol acyltransferase macromolecule monomer novel novel strategies programs protein protein interaction protein structure function receptor receptor-mediated signaling reconstitution single molecule transmission process

项目摘要

The mechanism underlying G protein-coupled receptor (GPCR)-mediated signaling systems is still unresolved despite the intense focus these systems have received over the past century. This is due in part to the difficulties in studying membrane proteins in their native context by methods that provide molecular details. The focus of the current proposal is to apply novel biophysical methodologies to unravel the molecular and temporal mysteries of GPCR-mediated signaling pathways. Rhodopsin and the visual system will be the initial focus of the research program. This prototypical GPCR signaling system offers several advantages that will allow for the application of novel biophysical approaches. Atomic force microscopy will result in high-resolution images of individual molecules that will provide structural and organizational information of the system. Single-molecule force spectroscopy will provide detailed information on the molecular interactions in rhodopsin that stabilize the protein and promote its function. Cryo-electron tomography will gain access to an unperturbed rod outer segment to provide structural information on this compartment and on the macromolecules that carry out their function at this venue. Fluorescence resonance energy transfer will be utilized to detect protein-protein interactions of signaling proteins to monitor the dynamic interactions that define the signaling process and the timeframe in which this takes place. Together the information obtained by this unique combination of methodologies will provide key pieces of molecular information that is currently unavailable for these systems. This will help define the molecular mechanism underlying the signaling events that govern important physiological processes regulated by GPCRs. G protein-coupled receptors (GPCRs) represent the largest class of cell surface proteins and drug targets currently on the market. This family of proteins is involved in virtually every physiological process, and dysfunctions in these systems can lead to diseases such as blindness, addiction, diabetes, and heart disease. Despite the importance of GPCRs an accurate molecular description of their action is still lacking. Understanding the molecular mysteries of these systems will lead to the development of more effective therapeutic solutions.

G蛋白偶联受体（GPCR）介导的信号系统的潜在机制仍然是尽管这些系统在过去一个世纪中受到了强烈关注，但仍未得到解决。这部分是由于解决了通过提供分子方法来研究膜蛋白在其天然环境中的困难细节。当前提案的重点是应用新颖的生物物理方法来解开 GPCR 介导的信号通路的分子和时间奥秘。视紫红质和视觉系统将是该研究计划的最初重点。这个原型 GPCR 信号系统提供了多种的优势将允许应用新颖的生物物理方法。原子力显微镜将产生单个分子的高分辨率图像，从而提供结构和组织系统的信息。单分子力谱将提供有关视紫红质中的分子相互作用可稳定蛋白质并促进其功能。冷冻电子断层扫描将获得未受干扰的杆外段，以提供有关该杆的结构信息隔室和在该场所发挥其功能的大分子。荧光共振能量转移将用于检测信号蛋白的蛋白质-蛋白质相互作用，以监测定义信号发送过程和发生时间范围的动态交互。一起通过这种独特的方法组合获得的信息将提供分子生物学的关键部分这些系统当前不可用的信息。这将有助于定义分子机制是控制 GPCR 调节的重要生理过程的信号事件的基础。 G 蛋白偶联受体 (GPCR) 代表最大类别的细胞表面蛋白和药物靶点目前市场上。该蛋白质家族几乎参与每一个生理过程，并且这些系统的功能障碍可能导致失明、成瘾、糖尿病和心脏病等疾病疾病。尽管 GPCR 很重要，但仍然缺乏对其作用的准确分子描述。了解这些系统的分子奥秘将有助于开发更有效的系统治疗解决方案。