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: 8505499
• 负责人: RUBEN ABAGYAN
• 金额: $ 86.69万
• 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/8505499
• 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 Targeting; Electron Spin Resonance Spectroscopy; Emotions; Endocrine system; 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.

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，通过转导涉及感觉、神经传递、发育、情绪、认知以及中枢神经系统、内分泌和免疫系统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，以其在免疫监视中的关键作用而闻名，它们控制白细胞的迁移和激活，以检测和解决癌症和感染等生理异常。然而，这些受体的不适当表达或调节与大量病理学相关，包括炎症性疾病、癌症和艾滋病；因此，开发阻断特定趋化因子受体功能的小分子受体拮抗剂引起了人们的极大兴趣。直到最近，由于受体表达和结晶方面的挑战，GPCR 仍无法确定结构。然而，新技术的出现使得确定 GPCR 结构的可行性无可争议。为此，我们的主要目标是获得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，申请 用于表达、纯化和结晶趋化因子受体家族 GPCR 的新技术，目标是在五年终点时确定至少两种不同的受体结构和多种复合物。为了最大限度地发挥 PSI 中心生成纯化蛋白质的能力，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辅相成。将开发并应用放射分解足迹来绘制趋化因子和受体之间的结合界面，并确定有关激活机制的信息。致病蛋白与趋化因子受体的相互作用也将被研究，特别是 CCR5 受体与 HIV 糖蛋白 gp120 的相互作用，以及 DARC 受体与疟疾对接蛋白 DBP 的相互作用。电子顺磁共振定点自旋标记 (SDSL-EPR) 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得到增强，以便合理指导实验结构设计并在 3D 背景下解释生物物理数据。