Structure-function studies of the membrane-interacting domains of HIV-1 Env spike

HIV-1 Env 刺突膜相互作用域的结构功能研究

• 批准号: 9203214
• 负责人: JAMES Jeiwen CHOU
• 金额: $ 85.83万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-20 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9203214
• 关键词: Adopted; Alanine; Antigens; Arginine; Binding; Biochemistry; Biogenesis; Biological Assay; C-terminal; CCR5 gene; CD4 Antigens; CXCR4 gene; Cell fusion; Cell membrane; Cell surface; Cells; Charge; Cleaved cell; Clinical Research; Computer Simulation; Coupling; Crystallography; Cytoplasmic Tail; Data; Development; Elements; Environment; Epitopes; Event; Goals; HIV; HIV Envelope Protein gp120; HIV-1; HIV-1 vaccine; HIV-2; Infection; Knowledge; Lead; Lipid Bilayers; Lipids; Lysine; Mediating; Membrane; Membrane Fusion; Molecular Conformation; Mutagenesis; Mutation; Peptides; Play; Production; Property; Protein Engineering; Proteins; Resolution; Role; SIV; Scanning; Side; Structure; Surface; Technology; Testing; Transmembrane Domain; Viral; Virion; Virus Assembly; Virus Diseases; base; design; env Glycoproteins; gp160; insight; large scale production; nanodisk; neutralizing antibody; novel; receptor; reconstitution; structural biology; vaccine development

Project Summary The first critical step of HIV-1 infection is fusion of viral and target cell membranes mediated by envelope glycoprotein (Env). The mature Env spikes [trimeric (gp160)3, cleaved to (gp120/gp41)3] are the sole antigens on the virion surface. Conformational changes in gp120 when triggered by binding to receptor (CD4) and co- receptor (e.g., CCR5 or CXCR4) lead to a cascade of refolding events in gp41, and ultimately to membrane fusion. Vast amount of structural information is available for the ectodomain of Env, but much less is known regarding its transmembrane domain (TMD) and its membrane-proximal (MP) regions, including the membrane proximal external region (MPER) and the cytoplasmic tail (CT). We recently made an unexpected discovery that truncation of the CT domain drastically reshapes the antigenic surfaces of the Env ectodomain on the other side of the membrane. Deep understanding of the physical coupling (conformation and/or dynamics) between the CT and the ectodomain mediated by the TMD may guide Env-based immunogen design to induce broadly neutralizing antibodies (bnNabs). We thus hypothesize that the membrane-interacting domains (MPER, TMD and CT) of HIV-1 Env adopt defined structures which are critical for its stability, function and antigenicity. We have already completed a TMD structure at the atomic resolution using the state-of-the- art NMR technology. When reconstituted in bicelles that mimic lipid bilayer, the TMD forms a well-ordered trimer mainly stabilized by a tightly packed hydrophilic core near its C-terminal end that can potentially be influenced by the CT domain. In this application, we plan to combine structural biology approaches and functional assays to elucidate the roles of the membrane-interacting domains of HIV-1 Env. We will purse the following specific aims: 1) we will investigate TMD assembly of HIV and SIV Env and possible interaction with the fusion peptide; 2) we will provide structural information for the MP regions of Env in the context of membrane; 3) we will elucidate the role of the membrane-interacting domains of Env in its stability, function and antigenicity; 4) we will design trimer immunogens to mimic the native and functional HIV-1 Env spike..

项目概要 HIV-1感染的第一个关键步骤是由包膜介导的病毒与靶细胞膜的融合 糖蛋白（Env）。成熟的 Env 刺突 [三聚体 (gp160)3，裂解为 (gp120/gp41)3] 是唯一的抗原 在病毒颗粒表面。与受体 (CD4) 和 co- 结合触发时 gp120 的构象变化 受体（例如 CCR5 或 CXCR4）导致 gp41 中的一系列重折叠事件，并最终导致膜 融合。 Env 胞外域有大量结构信息，但知之甚少 关于其跨膜结构域（TMD）和近膜（MP）区域，包括膜 近端外部区域（MPER）和细胞质尾部（CT）。最近我们有一个意想不到的发现 CT结构域的截断极大地重塑了Env胞外域的抗原表面 膜的另一面。对物理耦合（构象和/或动力学）的深入理解 CT 和 TMD 介导的胞外域之间的相互作用可能会指导基于 Env 的免疫原设计来诱导 广泛中和抗体（bnNabs）。因此，我们假设膜相互作用域 HIV-1 Env（MPER、TMD 和 CT）采用对其稳定性、功能至关重要的明确结构 和抗原性。我们已经使用最新技术完成了原子分辨率的 TMD 结构 艺术核磁共振技术。当在模拟脂质双层的 bicelles 中重建时，TMD 形成一个有序的结构 三聚体主要由 C 末端附近紧密堆积的亲水核心稳定，该核心可能被 受 CT 域的影响。在此应用中，我们计划结合结构生物学方法和 功能测定阐明 HIV-1 Env 膜相互作用域的作用。我们将钱包 以下具体目标：1）我们将研究 HIV 和 SIV Env 的 TMD 组装以及可能的相互作用 融合肽； 2）我们将在以下背景下提供 Env 的 MP 区域的结构信息 膜; 3）我们将阐明Env的膜相互作用域在其稳定性、功能中的作用 和抗原性； 4) 我们将设计三聚体免疫原来模拟天然和功能性的 HIV-1 Env 刺突。