STRUCTURAL BASIS OF CHEMOKINE RECEPTOR RECOGNITION

趋化因子受体识别的结构基础

• 批准号: 6372710
• 负责人: MICHAEL E HODSDON
• 金额: $ 12.8万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6372710
• 关键词: G protein; chemokine; circular dichroism; conformation; cytokine receptors; intermolecular interaction; laboratory mouse; macrophage inflammatory proteins; nuclear magnetic resonance spectroscopy; physical model; protein structure function; receptor binding; receptor expression; structural biology; synthetic peptide

The biological function of chemokines is critically dependent on their recognition and activation of specific G protein coupled receptors (GPCRs). The overall goal of the proposed project is to develop an understanding of the structural interactions between chemokines and their receptors. Unfortunately, these interactions cannot be directly observed by X-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy due to the difficulty of working with full-length GPCRs. Mutagenesis studies have identified residues that are involved in functional protein-protein interactions, but the precise interactions that result in receptor recognition and activation remain uncharacterized. The approach adopted in the proposed research attempts to circumvent this problem by reconstructing the recognition interface using synthetic peptides derived from the extracellular domains of chemokine receptors. The conformation of these receptor-based peptides and their interactions with chemokines in solution will then be characterized using circular dichroism and NMR spectroscopy. The biological activity of the peptides will be determined using in vitro assays of neutrophil chemotaxis and receptor binding. The data will be analyzed in order to develop a structural model of chemokine receptor recognition which will be tested using site-directed mutagenesis, followed by functional assays. Research into the structural basis of chemokine receptor recognition has wide-ranging relevance to human health and disease. Chemokines play a predominant role in a variety of allergic and rheumatic diseases including asthma, arthritis and psoriasis. Effects of chemokines on atherogenesis, angiogenesis and tumor growth have been reported. Some of the chemokine receptors function as HIV-1 coreceptors and have become attractive targets for identifying new anti-HIV compounds. A model of receptor recognition will guide experiments to delineate the biological roles of specific chemokine-receptor interactions. As well, therapeutic agents could be rationally designed to precisely inhibit single pairs of chemokines and their receptors which are specifically implicated in disease. The proposed project to be performed in the laboratory of Dr. Elias Lolis will allow me to continue my training in protein structural biology as well as introduce me to the science and techniques of immunobiology. My career development will be enhanced by this training and will prepare me for a future career in academic medical research.

趋化因子的生物学功能主要依赖于它们对特异性G蛋白偶联受体（GPCR）的识别和激活。 该项目的总体目标是了解趋化因子及其受体之间的结构相互作用。 不幸的是，由于难以使用全长GPCR，这些相互作用不能通过X射线晶体学或核磁共振（NMR）光谱直接观察到。 突变研究已经确定了参与功能性蛋白质-蛋白质相互作用的残基，但导致受体识别和激活的精确相互作用仍然没有表征。 在拟议的研究中采用的方法试图绕过这个问题，重建识别界面，使用合成肽衍生自细胞外结构域的趋化因子受体。 这些受体为基础的肽的构象和它们在溶液中与趋化因子的相互作用，然后将其特征在于使用圆二色谱和NMR光谱。 将使用嗜中性粒细胞趋化性和受体结合的体外测定来确定肽的生物活性。 将分析数据以开发趋化因子受体识别的结构模型，其将使用定点诱变进行测试，然后进行功能测定。对趋化因子受体识别的结构基础的研究与人类健康和疾病具有广泛的相关性。 趋化因子在包括哮喘、关节炎和牛皮癣在内的多种过敏性和风湿性疾病中起主要作用。 趋化因子在动脉粥样硬化形成、血管生成和肿瘤生长中的作用已有报道。 一些趋化因子受体作为HIV-1辅助受体发挥作用，并已成为鉴定新的抗HIV化合物的有吸引力的靶点。 受体识别的模型将指导实验来描述特定趋化因子-受体相互作用的生物学作用。同样，可以合理地设计治疗剂以精确地抑制与疾病特异性相关的单对趋化因子及其受体。在埃利亚斯·洛利斯博士的实验室进行的拟议项目将使我能够继续接受蛋白质结构生物学的培训，并向我介绍免疫生物学的科学和技术。 我的职业发展将通过这次培训得到加强，并将为我未来的学术医学研究生涯做好准备。