Structural Basis of Coreceptor Recognition by HIV-1 Envelope Spike

HIV-1 包膜尖峰识别辅助受体的结构基础

• 批准号: 10390469
• 负责人: Bing Chen
• 金额: $ 52.18万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-15 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390469
• 关键词: Address; Affinity; Architecture; Binding; Biological Assay; CCR5 gene; CD4 Antigens; CXCR4 gene; Cell membrane; Cells; Complex; Complex Analysis; Cryoelectron Microscopy; Crystallization; Data; Data Set; Detergents; Disease; Electron Microscopy; Elements; Embryo; Event; Family; G-Protein-Coupled Receptors; Glycoproteins; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Human; Infection; Kidney; Knowledge; Ligands; Lipid Bilayers; Membrane Fusion; Membrane Proteins; Microscope; Molecular; Mutagenesis; Mutation; Process; Reporting; Resolution; Role; Structure; Time; Viral; Virus; Virus Diseases; Work; chemokine; chemokine receptor; chronic infection; design; falls; gp160; molecular modeling; nanodisk; receptor; receptor function; small molecule; structural biology; transmission process

Project Summary HIV infection begins with fusion of viral and target cell membranes, driven by the viral envelope glycoprotein [Env; trimeric (gp160)3 cleaved to (gp120/gp41)3]. Gp120 binding to primary receptor CD4 and coreceptor (e.g. chemokine receptor CCR5 or CXCR4) trigger large structural rearrangements in gp41 that drive the membrane fusion process. Encounter of the coreceptor by gp120 is believed to be the crucial trigger for gp41 refolding events, which promotes membrane fusion. It has been two decades since CCR5 and CXCR4 were first identified as the coreceptors for HIV-1 entry, but we still do not have a clear picture, in particular, at atomic resolution, of how the coreceptor recognizes HIV-1 Env, except for some speculative molecular modeling. The coreceptors are chemokine receptors with seven transmembrane-spanning segments (7TMs) and belong to the family of G protein-coupled receptors (GPCRs). Crystal structures have been reported for heavily modified CCR5 and CXCR4, revealing the general architecture of these receptors, as well as their interactions with the various ligands, but the structures fall short of explaining the molecular details of how these chemokine receptors function as HIV-1 coreceptors. In this proposal, we plan to gain a better understanding of the HIV-1 coreceptor function of CCR5 and CXCR4 and to provide high-resolution pictures of how they interact with HIV- 1 Env to promote viral entry. We hypothesize that an extensive interface between gp120 and the coreceptor involving multiple structural elements is required for their high-affinity interaction. We have already purified a stable complex of HIV-1 gp120, 4 domain CD4 and an unmodified human CCR5, and demonstrated the feasibility to carry out electron microscopy (EM) studies. Recent advances in cryo-electron microscopy (cryoEM) have revolutionized the field of structural biology and produced numerous high-resolution structures. To capitalize on these advances, we will tackle a challenging problem that is important to both the HIV and GPCR fields. We will pursue following specific aims: 1) we will determine the high-resolution structure of the complex of CD4-gp120-CCR5 by cryoEM; 2) we will determine the high-resolution structure of the complex of CD4-gp120-CXCR4; 3) we will elucidate the role of key structural elements of CCR5 or CXCR4 in coreceptor function by structure-guided mutagenesis.

项目摘要 HIV感染始于病毒和靶细胞膜的融合，由病毒膜糖蛋白驱动 [env;三聚体（GP160）3裂解至（gp120/gp41）3]。 gp120与一级受体CD4和cocector结合（例如 趋化因子受体CCR5或CXCR4）在GP41中触发大型结构重排，该重排驱动膜 融合过程。据信，gp120遇到的cocector是GP41重折叠的关键触发因素 事件，促进膜融合。自CCR5和CXCR4首先以来已经二十年了 被识别为HIV-1进入的共感受器，但我们仍然没有明确的图片，特别是在原子上 除了某些投机性分子建模外，共素受体如何识别HIV-1 Env。这 共感受器是趋化因子受体，具有七个跨膜跨膜段（7TMS），属于 G蛋白偶联受体（GPCR）家族。据报道，晶体结构已进行了重大修饰 CCR5和CXCR4，揭示了这些受体的一般结构，以及它们与 各种配体，但结构尚未解释这些趋化因子的分子细节 受体充当HIV-1共感受器。在此提案中，我们计划更好地了解HIV-1 CCR5和CXCR4的croecector功能，并提供高分辨率的图片，说明它们如何与HIV相互作用 1 env促进病毒进入。我们假设GP120和 涉及多个结构元素的共核能是其高亲和力相互作用所必需的。我们有 已经纯化了HIV-1 GP120、4个域CD4和一个未修饰的人CCR5的稳定复合物，并且 证明了进行电子显微镜（EM）研究的可行性。冷冻电子的最新进展 显微镜（冷冻）彻底改变了结构生物学领域，并产生了许多高分辨率 结构。为了利用这些进步，我们将解决一个具有挑战性的问题，这对既重要 艾滋病毒和GPCR领域。我们将遵循特定目标：1）我们将确定高分辨率结构 Cryoem的CD4-GP120-CCR5复合物； 2）我们将确定该高分辨率的结构 CD4-GP120-CXCR4的复合物； 3）我们将阐明CCR5或CXCR4关键结构元素在 结构引导的诱变作用。

项目成果

Structural basis for enhanced infectivity and immune evasion of SARS-CoV-2 variants.

• DOI: 10.1126/science.abi9745
• 发表时间: 2021-08-06
• 期刊: SCIENCE
• 影响因子: 56.9
• 作者: Cai, Yongfei;Zhang, Jun;Xiao, Tianshu;Lavine, Christy L.;Rawson, Shaun;Peng, Hanqin;Zhu, Haisun;Anand, Krishna;Tong, Pei;Gautam, Avneesh;Lu, Shen;Sterling, Sarah M.;Walsh, Richard M.;Rits-Volloch, Sophia;Lu, Jianming;Wesemann, Duane R.;Yang, Wei;Seaman, Michael S.;Chen, Bing
• 通讯作者: Chen, Bing

HIV-1 fusion inhibitors targeting the membrane-proximal external region of Env spikes.

• DOI: 10.1038/s41589-020-0496-y
• 发表时间: 2020-05
• 期刊: Nature chemical biology
• 影响因子: 14.8
• 作者: Xiao T;Frey G;Fu Q;Lavine CL;Scott DA;Seaman MS;Chou JJ;Chen B
• 通讯作者: Chen B