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：
7692473
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/7692473
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 Class Complex Condition Cyan Fluorescent Protein Cyclophosphamide/Fluorouracil/Prednisone DNA Sequence Rearrangement Data Detection Development Diabetes Mellitus Dimensions Disease Dithiothreitol Drug Delivery Systems Electron Microscopy Electrophysiology (science)Energy Transfer Ethylmaleimide Event Exhibits Fluorescence Resonance Energy Transfer Fluorescence Spectroscopy Foundations Functional disorder G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors Genes Genetic Green Fluorescent Proteins Heart Diseases Helix (Snails)Housing Image Individual Information Systems Invasive Investigation Ions Knowledge Lead Life Light Lipids Location Maintenance Marketing Measures Mediating Membrane Membrane Proteins Mentors Methodology Methods Microscopy Modification Molecular Molecular Biology Monitor Movement Nature Noise Optical Methods Organelles Pathway interactions Phase Phototransduction Physiological Processes Play Process Property Protein Family Proteins Purpose 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 ear helix 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 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）介导的信号系统的基础机制仍然是尽管这些系统在过去的一个世纪中获得了强烈的重点，但仍未解决。这部分应通过提供分子的方法，在本地情况下研究膜蛋白的困难细节。当前建议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谜。视紫红素和视觉系统将是研究计划的最初重点。这个典型的GPCR信号系统提供了多个将允许应用新型生物物理方法的优势。原子力显微镜将导致单个分子的高分辨率图像，这些图像将提供结构和组织系统的信息。单分子力光谱将提供有关该的详细信息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不受干扰的杆外部段，以提供有关此的结构信息在此场地执行其功能的大分子隔间和大分子上。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过程和发生时间范围的动态交互。一起这种独特的方法组合获得的信息将提供关键的分子这些系统目前无法使用的信息。这将有助于定义分子机制基本的信号事件，该信号事件控制了由GPCR调节的重要生理过程。 G蛋白偶联受体（GPCR）代表最大的细胞表面蛋白和药物靶标目前在市场上。这个蛋白质家族几乎参与了每个生理过程，这些系统的功能障碍会导致失明，成瘾，糖尿病和心脏等疾病疾病。尽管GPCR的重要性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导致更有效的发展治疗解决方案。