Investigating the Architecture of HIV Envelope Glycoprotein

研究 HIV 包膜糖蛋白的结构

• 批准号: 8262992
• 负责人: Miklos Guttman
• 金额: $ 4.92万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8262992
• 关键词: Address; Antibodies; Antigens; Architecture; Binding; Binding Sites; CD4 Antigens; Carbohydrates; Cataloging; Catalogs; Cells; Comparative Study; Complementarity Determining Regions; Complex; Coupled; Data; Deuterium; Development; Enzyme-Linked Immunosorbent Assay; Epitopes; Glycoproteins; Goals; HIV; HIV Envelope Protein gp120; Hydrogen; Imagery; Immune; Immune response; Immunoprecipitation; Kinetics; Link; Maps; Masks; Mass Spectrum Analysis; Measures; Mediating; Membrane; Membrane Proteins; Methods; Modeling; Molecular; Monitor; Monoclonal Antibodies; Nature; Polysaccharides; Property; Proteins; Resolution; Roentgen Rays; Role; Shapes; Site; Solvents; Spatial Distribution; Structure; System; Techniques; Testing; Uncertainty; Vaccine Design; Vaccines; Viral; base; design; env Glycoproteins; flexibility; glycosylation; gp160; improved; interest; neutralizing antibody; next generation; receptor; receptor binding; reconstruction; vaccine development; virus envelope

DESCRIPTION (provided by applicant): The human immunodeficiency virus (HIV) employs an array of properties that enable it to evade antibody neutralization. Rational vaccine design has been limited due to the insufficient structural understanding of the HIV envelope glycoprotein (Env), the primary target for neutralizing antibodies. Advances in structural determination have been hindered due to the presence of flexible hypervariable loops and the high carbohydrate content of this protein and the arrangement of the full Env complex remains controversial. The nature of the glycan shield, thought to mask conserved sites, is also of key interest as it mediates immune escape. Furthermore the binding of host cell CD4 receptor is known to induce massive conformational changes in Env, but a detailed examination of these changes has yet to be addressed. This proposal aims to combine two complementary approaches, small-angle X-ray scattering (SAXS) with ab initio shape reconstruction and hydrogen-deuterium exchange (HXMS) with mass spectrometry analysis to address several questions regarding the architecture of trimeric Env. The extensive catalog of constructs available from the Hu lab enables comparative studies of Env ranging from monomeric subunits to trimeric complexes, in the context of the same isolate. Identification of the trimeric interface, conformational changes induced upon receptor binding, and epitope occlusion by variable loops and glycans can be addressed with these techniques. The interaction of several well-characterized monoclonal antibodies directed against the CD4 binding site will also be compared to address the question of how conformational changes induced by antibody binding differ from those resulting from CD4 binding, and whether there are potentially correlates with neutralizing potency. Understanding these properties of Env will assist current efforts to identify targets for development of broadly neutralizing antibodies, necessary for an effective vaccine. PUBLIC HEALTH RELEVANCE: The poor understanding of the primary surface protein of the Human Immunodeficiency Virus (HIV) has been a major hindrance to vaccine development. A thorough understanding of the organization of this protein is critical for designing next generation immunogens to induce an effective immune response. Using two emerging techniques we will address uncertainties regarding the architecture of this surface protein, identify how critical antigenic regions are protected, and identify ways to improve vaccine design.

描述（由申请人提供）：人类免疫缺陷病毒（HIV）采用了一系列特性，使其能够逃避抗体中和。由于对HIV包膜糖蛋白（ENV）的结构理解不足，这是中和抗体中和抗体的主要靶标，理性疫苗的设计受到限制。由于存在柔性高变量环和该蛋白质的高碳水化合物含量，因此结构测定的进展受到阻碍。被认为是掩盖保守的地点的聚糖盾牌的性质也具有关键的兴趣，因为它介导了免疫逃生。此外，已知宿主细胞CD4受体的结合会引起ENV中的巨大构象变化，但是对这些变化的详细检查尚未解决。 该建议旨在将两种互补方法，小角度的X射线散射（SAX）与从头开始形状重建和氢 - 偏见交换（HXMS）和质谱分析结合在一起，以解决有关Trimeric Env的结构的几个问题。在同一分离株的背景下，HU实验室可用的广泛的构造目录可以对从单体亚基到三聚体络合物的比较研究。可以用这些技术来识别三聚体界面，受体结合对受体结合引起的构象变化以及通过可变环和聚糖的表位闭塞。还将比较几种针对CD4结合位点的几种特征良好的单克隆抗体的相互作用，以解决抗体结合引起的构象变化与CD4结合引起的构象变化的问题，并且是否存在潜在的与中和效力相关的问题。了解ENV的这些特性将有助于当前的努力，以确定开发有效疫苗所必需的广泛中和抗体的目标。 公共卫生相关性：对人类免疫缺陷病毒（HIV）原发性表面蛋白（HIV）的不良理解一直是疫苗开发的主要障碍。对该蛋白质的组织的透彻理解对于设计下一代免疫原以诱导有效的免疫反应至关重要。使用两种新兴技术，我们将解决有关该表面蛋白结构的不确定性，确定如何保护关键的抗原区域，并确定改善疫苗设计的方法。