Structure, Dynamics and Activation Mechanisms of Chemokine Receptors

趋化因子受体的结构、动力学和激活机制

基本信息

批准号：
7982295
负责人：
RUBEN ABAGYAN
金额：
$ 100.81万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-30 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7982295
关键词：
Acquired Immunodeficiency Syndrome Affinity Agonist Arrestins Asthma Binding Biological Process Biology Cells Chemokine (C-C Motif) Receptor 5 Cocrystallography Cognition Collaborations Complement Complex Computer Analysis Computer Simulation Coupling Crystallization Crystallography Data Deposition Development Disease Docking Drug Delivery Systems Electron Spin Resonance Spectroscopy Emotions Endocrine Ensure Esthesia Event Family G-Protein-Coupled Receptors GTP-Binding Proteins Glycoproteins Goals HIV HIV Envelope Protein gp120 Heart Diseases Human Immune system Immunologic Surveillance Infection Inflammatory Insecta Internet Intracellular Signaling Proteins Ligand Binding Ligands Light Malignant Neoplasms Mammalian Cell Maps Mass Spectrum Analysis Membrane Methodology Methods Modeling Modification Multiple Sclerosis Mutagenesis Mutation Output Pathology Pharmaceutical Preparations Physiological Play Principal Investigator Production Property Protein Structure Initiative Proteins Protocols documentation Receptor Activation Regulation Resolution Rheumatoid Arthritis Role Signal Transduction Signaling Protein Site Solvents Source Spin Labels Stimulus Structural Models Structure Surface Synchrotrons Technology Work base chemokine chemokine receptor data integration design design and construction drug discovery human disease improved innovation insight intercellular communication interest knowledge base leukocyte activation member migration mutant neurotransmission new technology overexpression public health relevance receptor receptor binding receptor expression receptor function restraint small molecule structural genomics

项目摘要

DESCRIPTION (provided by applicant): G protein coupled receptors (GPCRs) represent the largest protein superfamily in humans, with nearly 1000 members. They are seven transmembrane receptors that coordinate intercellular communication via the transduction of a wide range of stimuli involved in sensation, neurotransmission, development, emotion, cognition, and function in the CNS, endocrine and immune systems. Chemokine receptors are an important class of GPCRs that are best known for their pivotal role in immune surveillance, where they control the migration and activation of leukocytes in an effort to detect and resolve physiological abnormalities such as cancer and infection. However, inappropriate expression or regulation of these receptors is associated with an extraordinary number of pathologies including inflammatory diseases, cancer and AIDS; thus there is significant interest in developing small molecule receptor antagonists that block the function of specific chemokine receptors. Until recently, GPCRs had eluded structure determination due to challenges in receptor expression and crystallization. However, new technologies have emerged which has made the viability of determining GPCR structures indisputable. To this end, our primary goal is to obtain structural information on chemokine receptors and receptor complexes that can aid drug discovery efforts aimed at improving affinity, efficacy, and selectivity. Accordingly, in collaboration with the PSI network, we will apply novel technologies for the expression, purification and crystallization of GPCRs from the chemokine receptor family, with the goal of determining at least two different receptor structures and multiple co-complexes by the five-year endpoint. To maximize the capabilities of the PSI centers in generating purified protein, and to acquire insights into the dynamic aspects of receptor function, the crystallographic work will be complemented with biophysical studies. Radiolytic footprinting will be developed and applied to map the binding interfaces between chemokines and receptors and to determine information on activation mechanisms. The interaction of pathogenic proteins with chemokine receptors will also be investigated, specifically, the CCR5 receptor with the HIV glycoprotein gp120, and the DARC receptor with the malarial docking protein, DBP. Site Directed Spin Labeling with Electron Paramagnetic Resonance (SDSL-EPR) will be used to characterize the conformational changes associated with ligand binding. All of these studies will be augmented with computational modeling methods in order to rationally guide the experimental construct design and to interpret the biophysical data in a 3D context. PUBLIC HEALTH RELEVANCE: Chemokine receptors are involved in many human diseases including asthma, multiple sclerosis, rheumatoid arthritis, heart disease, cancer and HIV, Determining structures of chemokine receptors in complex with small molecule drugs and ligands, and information on ligand-induced conformational changes, will significantly aid the development of drugs targeting these receptors.

描述（由申请人提供）：G蛋白偶联受体（GPCR）代表了人类中最大的蛋白质超家族，近1000名成员。它们是七个跨膜受体，通过转导参与中枢神经系统，内分泌和免疫系统中的感觉，神经传递，发育，情感，认知和功能的广泛刺激来协调细胞间通信。趋化因子受体是一类重要类别的GPCR，以其在免疫监测中的关键作用而闻名，它们控制着白细胞的迁移和激活，以检测和解决生理异常，例如癌症和感染。但是，这些受体的不当表达或调节与包括炎症性疾病，癌症和艾滋病在内的大量病理有关。因此，人们对开发阻断特定趋化因子受体功能的小分子受体拮抗剂的兴趣很大。直到最近，由于受体表达和结晶的挑战，GPCR还避免了结构测定。但是，已经出现了新技术，这使确定GPCR结构无可争议的生存能力。为此，我们的主要目标是获取有关趋化因子受体和受体复合物的结构信息，这些信息可以帮助旨在提高亲和力，功效和选择性的药物发现工作。因此，与PSI网络合作，我们将申请从趋化因子受体家族中GPCR的表达，纯化和结晶的新技术，目的是通过五年端点确定至少两个不同的受体结构和多个共络合物。为了最大程度地提高PSI中心产生纯化蛋白质的能力，并获得对受体功能动态方面的见解，晶体学工作将与生物物理研究相辅相成。放射性足迹将开发并应用于绘制趋化因子和受体之间的结合界面，并确定激活机制的信息。病原蛋白与趋化因子受体的相互作用也将被研究，具体来说，CCR5受体与HIV糖蛋白GP120和具有疟疾对接蛋白DBP的DARC受体DBP。用电子顺磁共振（SDSL-EPR）定向自旋标记将用于表征与配体结合相关的构象变化。所有这些研究都将通过计算建模方法进行增强，以便在3D环境中合理地指导实验构造设计并解释生物物理数据。公共卫生相关性：趋化因子受体参与许多人类疾病，包括哮喘，多发性硬化症，类风湿关节炎，心脏病，癌症和HIV，确定与小分子药物和配体复合物中趋化因子受体的结构，以及有关配体诱导的构象变化的信息，将有助于靶向这些受体的发育。