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..

项目摘要 HIV-1感染的第一个关键步骤是由包膜介导的病毒和靶细胞膜融合 糖蛋白（Env）。成熟的Env Spikes [Trimeric（GP160）3，裂解至（GP120/GP41）3]是唯一的抗原 在病毒体表面。当通过与受体（CD4）和共同的结合触发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，最终导致膜 融合。 ENV的外生域可用大量的结构信息，但少得多 关于其跨膜结构域（TMD）及其膜 - 螺旋（MP）区域，包括膜 近端外部区域（MPER）和细胞质尾部（CT）。我们最近做出了意想不到的发现 CT结构域的截断急剧重塑了Env Ectodomain的抗原表面 膜的另一侧。对物理耦合（构象和/或动态）的深刻理解 在TMD介导的CT和胞外域之间，可以指导基于ENV的免疫原设计以诱导 广泛中和抗体（BNNABS）。因此，我们假设膜相互作用域 HIV-1 env的（MPER，TMD和CT）采用定义的结构，这些结构对于其稳定性至关重要，功能 和抗原性。我们已经使用最新的原子分辨率完成了TMD结构 艺术NMR技术。当在模仿脂质双层的双丝中重构时，TMD构成了一个有序的 三聚体主要被其C末端附近紧密堆积的亲水芯稳定，可能是 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 功能分析以阐明HIV-1 Env的膜相互作用结构域的作用。我们将钱包 以下具体目的：1）我们将研究HIV和SIV Env的TMD组装，并可能与 融合肽； 2）我们将在ENV的MP地区提供结构信息 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以模仿天然和功能性HIV-1 envike。