Quantitative analysis of metabotropic glutamate receptor activation and modulation

代谢型谷氨酸受体激活和调节的定量分析

基本信息

批准号：
10463681
负责人：
Reza Vafabakhsh
金额：
$ 33.25万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10463681
关键词：
Affect Allosteric Regulation Anxiety Architecture Binding Sites Biochemical Biological Biological Assay Biophysics Brain Calcium-Sensing Receptors Cell membrane Cells Chemicals Cholesterol Complement Complex Computing Methodologies Coupled Coupling Cysteine-Rich Domain Data Development Drug Targeting Environment Epilepsy Family Fluorescence Resonance Energy Transfer Fragile X Syndrome Functional disorder Funding G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors Glioma Glutamates Goals Grant Heterodimerization Human In Vitro Knowledge Learning Ligand Binding Ligands Lipids Malignant Neoplasms Mediating Membrane Membrane Proteins Metabotropic Glutamate Receptors Methodology Methods Microscopy Modeling Molecular Molecular Conformation Motion Mutagenesis Nature Parkinson Disease Pattern Pharmaceutical Preparations Physiological Process Property Protein Conformation Protein Dynamics Protein Engineering Protomer Psychotic Mood Disorders Receptor Activation Receptor Signaling Reporter Research Resolution Role Schizophrenia Sensory Shapes Signal Pathway Signal Transduction Signaling Molecule Site Spectrum Analysis Structure Techniques Time Tissues Transmembrane Domain Venus Flytrap Work addiction autism spectrum disorder base crosslink design dimer drug development glutamatergic signaling human disease insight intercellular communication live cell imaging member metabotropic glutamate receptor 2 millisecond multidisciplinary nanometer nervous system disorder neuroregulation new technology novel rational design receptor receptor function reconstitution side effect single molecule single-molecule FRET spectroscopic survey therapeutically effective

项目摘要

Project Summary G protein-coupled receptors (GPCRs) are the largest family of membrane receptors in human and have emerged as the largest family of drug targets with more than 35% of all drugs on the market functioning through GPCRs. Despite their desirability, to date, many GPCR families remain undrugged, partly due to lack of mechanistic knowledge about their activation and modulation. Metabotropic glutamate receptors (mGluRs) are members of class C GPCRs and are critical modulators of glutamate signaling. Due to their widespread expression in tissue and their central role they are among the most promising drug targets for the neurological disorders such as fragile X syndrome, epilepsy, anxiety, schizophrenia as well as some cancers. Advances in protein engineering and functional, structural and computational methods in the past twenty years have provided insights into the architecture, signaling and expression patterns of mGluRs. However, a general model of how ligands change the shape of mGluRs and how this conformational change is relayed across the membrane and over 12 nm to activate specific signaling pathways is poorly understood. In this research we will develop a novel technology that will allow us to watch a single mGluR protein while functioning in physiological conditions, in real time. This will allow us to quantify the motions of different domains of the receptor that are involved in signaling. Next, we will employ this approach to study how synthetic modulators of mGluR signaling affect protein conformation and dynamics to cholesterol and affect receptor signaling. This is a multi-disciplinary proposal where state-of-the-art in vitro single-molecule FRET (smFRET) spectroscopy is complemented by live-cell imaging, protein engineering and biophysical and biochemical methods. Once accomplished, the proposed research could provide a critical step towards rational design of efficient drugs with fewer undesirable side effects. Furthermore, these studies will provide a general roadmap for quantitative high-resolution structure-function studies of mammalian membrane proteins.

项目概要 G 蛋白偶联受体 (GPCR) 是人类最大的膜受体家族，具有成为最大的药物靶点家族，市场上超过 35% 的药物发挥作用通过 GPCR。尽管它们很受欢迎，但迄今为止，许多 GPCR 家族仍未被药物影响，部分原因是缺乏有关其激活和调节的机械知识。代谢型谷氨酸受体 (mGluR) 是 C 类 GPCR 的成员，是谷氨酸信号传导的关键调节剂。由于它们的广泛传播在组织中的表达及其核心作用它们是神经系统最有希望的药物靶标之一脆性 X 综合征、癫痫、焦虑症、精神分裂症以及某些癌症等疾病。进展过去二十年来蛋白质工程以及功能、结构和计算方法已经提供了对 mGluR 的结构、信号传导和表达模式的见解。然而，一般配体如何改变 mGluR 的形状以及这种构象变化如何在整个结构中传递的模型膜和超过 12 nm 激活特定信号传导途径的情况尚不清楚。在这项研究中我们将开发一项新技术，使我们能够在发挥作用时观察单个 mGluR 蛋白实时生理状况。这将使我们能够量化不同域的运动参与信号传导的受体。接下来，我们将采用这种方法来研究如何合成的调制器的谷氨酸受体信令影响蛋白质构象和动力学到胆固醇和影响受体发信号。这是一项多学科提案，其中采用了最先进的体外单分子 FRET (smFRET) 活细胞成像、蛋白质工程以及生物物理和生物化学对光谱学进行了补充方法。一旦完成，所提出的研究可以为合理设计提供关键的一步。有效的药物，不良副作用较少。此外，这些研究将提供一个总体路线图用于哺乳动物膜蛋白的定量高分辨率结构功能研究。