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.

项目概要 HIV 感染始于病毒包膜糖蛋白驱动的病毒与靶细胞膜的融合 [环境；三聚体 (gp160)3 裂解为 (gp120/gp41)3]。 Gp120 与主要受体 CD4 和辅助受体（例如 趋化因子受体 CCR5 或 CXCR4) 触发 gp41 中驱动膜的大结构重排 融合过程。 gp120 与辅助受体的相遇被认为是 gp41 重折叠的关键触发因素 事件，促进膜融合。自 CCR5 和 CXCR4 首次问世以来已有二十年了 被确定为 HIV-1 进入的辅助受体，但我们仍然没有清晰的认识，特别是在原子方面 分辨率，除了一些推测性分子模型外，辅助受体如何识别 HIV-1 Env。这 辅助受体是具有七个跨膜片段（7TM）的趋化因子受体，属于 G 蛋白偶联受体 (GPCR) 家族。据报道晶体结构已被严重修改 CCR5 和 CXCR4，揭示了这些受体的一般结构，以及它们与 各种配体，但结构不足以解释这些趋化因子如何产生的分子细节 受体作为 HIV-1 辅助受体发挥作用。在本提案中，我们计划更好地了解 HIV-1 CCR5 和 CXCR4 的辅助受体功能，并提供它们如何与 HIV 相互作用的高分辨率图片 1 Env 促进病毒进入。我们假设 gp120 和 涉及多个结构元件的共受体是它们的高亲和力相互作用所必需的。我们有 已经纯化了 HIV-1 gp120、4 个结构域 CD4 和未修饰的人类 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。低温电子学的最新进展 显微镜 (cryoEM) 彻底改变了结构生物学领域，并产生了大量高分辨率 结构。为了利用这些进步，我们将解决一个对双方都很重要的具有挑战性的问题 HIV 和 GPCR 领域。我们将追求以下具体目标：1）我们将确定高分辨率结构 通过冷冻电镜观察 CD4-gp120-CCR5 复合物； 2）我们将确定的高分辨率结构 CD4-gp120-CXCR4 复合物； 3）我们将阐明CCR5或CXCR4的关键结构元件在 通过结构引导诱变实现辅助受体功能。

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