Crystallographic studies of intact and fully glycosylated HIV-1 gp120

完整和完全糖基化的 HIV-1 gp120 的晶体学研究

• 批准号: 8603481
• 负责人: Bing Chen
• 金额: $ 41.26万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-20 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8603481
• 关键词: Antibodies; Antiviral Agents; Binding; Biochemical; CCR5 gene; CD4 Antigens; CXCR4 gene; Carbohydrates; Cell membrane; Cells; Characteristics; Chronic; Complex; Crystallization; DNA Sequence Rearrangement; Development; Evolution; Glycoproteins; HIV Envelope Protein gp120; HIV-1; Immune; Infection; Knowledge; Laboratories; Lead; Length; Ligands; Light; Mediating; Membrane Fusion; Molecular; Patients; Phase; Polysaccharides; Process; Protein Fragment; Proteins; Research; Resistance; Resolution; Solutions; Structure; Therapeutic; Tropism; Viral; Virus; base; chemokine receptor; env Gene Products; improved; insight; neutralizing antibody; novel; public health relevance; receptor; receptor binding; structural biology; therapeutic vaccine; vaccine development

DESCRIPTION (provided by applicant): HIV-1 infection begins with fusion of viral and target cell membranes. Viral attachment and membrane fusion are mediated by viral envelope spikes upon engagement with host cellular receptors. The mature envelope spikes contain three copies each of noncovalently-associated receptor-binding subunit gp120 and fusion subunit gp41. Despite considerable progress in our understanding of the structure of HIV-1 envelope glycoprotein over the last 15 years, we still do not have an atomic picture of the full-length and fully glycosylated gp120 due to enormous technical challenges associated with crystallographic studies of this protein, which is heavily coated with carbohydrates. Such a structure is, however, critical for a full understanding of gp120 function, as well as its interactions with various ligans, in particular, broadly neutralizing antibodies (bNAbs). Determination of an atomic structure of an intact HIV-1 gp120 will mark an important milestone in structural biology of HIV-1 entry, and may also guide development of antiviral therapeutics and vaccines. We have obtained diffracting crystals of a full-length and fully glycosylated HIV-1 gp120. In this application, we propose to determine crystal structures of the intact and glycosylated HIV-1 gp120 in complex with several neutralizing antibodies. We hypothesize that high-solution crystal structures of the unaltered HIV-1 gp120 will provide novel mechanistic insights into gp120 function, antibody neutralization and immune evasion. We will pursue the following specific aims: 1) We will determine the crystal structure of an intact, fully-glycosylated HIV-1 gp120; 2) We will determine the crystal structure of intact gp120 in complex with 2 domain CD4 and an anti-CD4 antibody ibalizumab; 3) We will determine crystal structures of HIV-1 gp120s with distinct characteristics.

描述（由申请人提供）：HIV-1 感染始于病毒和靶细胞膜的融合。病毒附着和膜融合是由病毒包膜刺突与宿主细胞受体结合介导的。成熟的包膜刺突包含非共价相关受体结合亚基 gp120 和融合亚基 gp41 各三个拷贝。尽管在过去 15 年里我们对 HIV-1 包膜糖蛋白结构的理解取得了相当大的进展，但由于这种蛋白质的晶体学研究面临着巨大的技术挑战，我们仍然没有全长和完全糖基化的 gp120 的原子图，该蛋白质被大量碳水化合物包裹。然而，这样的结构对于充分理解 gp120 功能及其与各种配体，特别是广泛中和抗体 (bNAb) 的相互作用至关重要。完整 HIV-1 gp120 原子结构的确定将标志着 HIV-1 进入结构生物学的一个重要里程碑，也可能指导抗病毒疗法和疫苗的开发。我们获得了全长且完全糖基化的 HIV-1 gp120 的衍射晶体。在此应用中，我们建议确定完整和糖基化的 HIV-1 gp120 与几种中和抗体复合物的晶体结构。我们假设未改变的 HIV-1 gp120 的高分辨晶体结构将为 gp120 功能、抗体中和和免疫逃避提供新的机制见解。我们将追求以下具体目标：1）我们将确定完整的、完全糖基化的HIV-1 gp120的晶体结构； 2) 我们将确定完整 gp120 与 2 个结构域 CD4 和抗 CD4 抗体 ibalizumab 复合物的晶体结构； 3）我们将确定具有独特特征的HIV-1 gp120的晶体结构。