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: 9053443
• 负责人: Bing Chen
• 金额: $ 44.25万
• 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/9053443
• 关键词: Antibodies; Antiviral Agents; Binding; Biochemical; CCR5 gene; CD4 Antigens; CXCR4 gene; Carbohydrates; Cell membrane; Cells; Characteristics; Chronic; Complex; Crystallization; Development; Evolution; Glycoproteins; HIV Envelope Protein gp120; HIV-1; Health; Immune; Infection; Knowledge; Laboratories; Lead; Length; Ligands; Light; Mediating; Membrane Fusion; Molecular; Patients; Phase; Polysaccharides; Process; Protein Fragment; Proteins; Research; Resistance; Resolution; Structure; Therapeutic; Tropism; Viral; Virus; base; chemokine receptor; env Gene Products; improved; insight; neutralizing antibody; novel; 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通过与宿主细胞受体互动后的病毒包膜尖峰介导的。成熟的包膜尖峰包含三个非共价相关受体结合亚基GP120和融合亚基GP41的副本。尽管在过去的15年中，我们对HIV-1包膜糖蛋白的结构的理解取得了长足的进步，但由于与该蛋白质的晶体学研究相关的巨大技术挑战，我们仍然没有对全长且完全糖基化的GP120的原子图，这与碳水化合物大致涂有碳水化合物。但是，这种结构对于对GP120功能的全面了解及其与各种Ligans的相互作用，特别是广泛中和抗体（BNAB）至关重要。确定完整的HIV-1 GP120原子结构将标志着HIV-1进入结构生物学的重要里程碑，还可以指导抗病毒疗法和疫苗的开发。我们获得了全长和完全糖基化的HIV-1 GP120的衍射晶体。在此应用中，我们建议在复合物中确定完整和糖基化的HIV-1 gp120的晶体结构，并具有几种中和抗体的晶体结构。我们假设未经改变的HIV-1 GP120的高解决晶体结构将为GP120功能，抗体中和和免疫逃避提供新颖的机械见解。我们将追求以下特定目标：1）我们将确定完整，完全糖基化的HIV-1 GP120的晶体结构； 2）我们将确定与2个结构域CD4和抗CD4抗体ibalizumab的复合物中完整的GP120的晶体结构； 3）我们将确定具有不同特征的HIV-1 GP120的晶体结构。