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

• 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
• 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

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）代表人类最大的蛋白质超家族，有近1000个成员。它们是七种跨膜受体，通过转导涉及CNS、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讯。 趋化因子受体是一类重要的GPCR，其最为人所知的是其在免疫监视中的关键作用，其中它们控制白细胞的迁移和活化，以检测和解决生理异常，如癌症和感染。然而，这些受体的不适当表达或调节与包括炎性疾病、癌症和AIDS在内的大量病理学相关;因此，人们对开发阻断特异性趋化因子受体功能的小分子受体拮抗剂非常感兴趣。直到最近，由于受体表达和结晶的挑战，GPCR一直无法确定结构。然而，新技术的出现使得确定气相化学还原结构的可行性无可争议。为此，我们的主要目标是获得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，我们将与PSI网络合作， 用于表达、纯化和结晶趋化因子受体家族GPCR的新技术，目标是在五年终点前确定至少两种不同的受体结构和多种共复合物。为了最大限度地提高PSI中心产生纯化蛋白质的能力，并深入了解受体功能的动态方面，晶体学工作将与生物物理学研究相结合。辐射分解足迹法将被开发和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。致病蛋白与趋化因子受体的相互作用也将被研究，特别是，CCR 5受体与HIV糖蛋白gp 120，和DARC受体与疟疾对接蛋白，DBP。将使用电子顺磁共振位点定向自旋标记（SDSL-EPR）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，以合理指导实验结构设计并在3D背景下解释生物物理数据。 公共卫生相关性：趋化因子受体与哮喘、多发性硬化症、类风湿性关节炎、心脏病、癌症和HIV等多种人类疾病有关。确定趋化因子受体与小分子药物和配体复合物的结构，以及配体诱导的构象变化的信息，将大大有助于开发靶向这些受体的药物。