HIV-1 and SIV Envelope Glycoproteins

HIV-1 和 SIV 包膜糖蛋白

• 批准号: 6866466
• 负责人: KENNETH H ROUX
• 金额: $ 32.41万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-02-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6866466
• 关键词: HIV envelope protein gp120; HIV envelope protein gp41; MHC class II antigen; X ray crystallography; computer simulation; cryoelectron microscopy; epitope mapping; human immunodeficiency virus 1; ligands; molecular site; monoclonal antibody; protein localization; protein purification; protein quantitation /detection; recombinant proteins; simian immunodeficiency virus; tomography; virion; virus envelope; virus morphology

DESCRIPTION (provided by applicant): Detailed knowledge of the structural and functional aspects of the molecules involved in the interaction, binding, and fusion of HIV-1 and the closely related SIV with their target cells is fundamental to our understanding of the disease processes in AIDS and to the rational design of diagnostic reagents, vaccines and other therapeutic agents. Two of the most important molecules in the infectious process, namely the envelope glycoproteins (Env) glycoproteins, SU (surface), and TM (transmembrane), are only partially characterized at the structural level with many of the important features are still poorly defined. These areas of uncertainty include the orientation of the variable loops, the structural changes that occur during the binding and fusion processes, and the features which foster or inhibit Ab-mediated neutralization. Another area of uncertainty involves the manner by which the Env proteins are expressed on the surface of virus particles. There is, for example, controversy regarding the number of Env spikes on the viral surface, their distribution and their stability. This proposal describes investigations aimed at applying new approaches for the determination of the 3-D morphology of the HIV-1 and SIV Env, gp120 and gp41, as expressed on the viral surface and in purified recombinant form. Our goals are to determine the numbers, locations, orientations, and structural details of Env molecules, both alone and in complex with monoclonal antibodies and ligands, on the surface of various forms of HIV-1 and SIV viral particles. In addition we will expand upon our previous epitope and reactive-site mapping studies of individual molecules and complexes by conducting 3-D structural analyses on 2-D crystals composed of purified monomeric and trimeric Env proteins and immune complexes. The primary methodologies will be the analysis of intact virions by negative stain and cryo-electron microscopic (EM) tomography and the analysis of purified recombinant Env and immune complexes by electron diffraction of 2-D crystalline arrays.

描述（由申请人提供）：详细了解HIV-1相互作用，结合和融合的分子的结构和功能方面以及与其靶细胞密切相关的SIV与我们对艾滋病中疾病过程以及诊断，疫苗，疫苗和其他治疗剂的合理设计的理解至关重要。传染过程中最重要的两个分子，即包络糖蛋白（ENV）糖蛋白，SU（表面）和TM（跨膜），仅在结构水平上部分表征，许多重要特征仍然不当定义。这些不确定性领域包括可变环的方向，结合和融合过程中发生的结构变化以及促进或抑制AB介导的中和的特征。不确定性的另一个区域涉及在病毒颗粒表面表达ENV蛋白的方式。例如，关于病毒表面上的Env峰值的数量，它们的分布和稳定性存在争议。该提案描述了旨在应用新方法来确定HIV-1和SIV Env，GP120和GP41的3-D形态，如在病毒表面和纯化的重组形式所示。我们的目标是在各种形式的HIV-1和SIV病毒颗粒的表面上，确定单独和与单克隆抗体和配体复杂的ENV分子的数量，位置，方向和结构细节。此外，我们将通过对由纯化的单体和三聚体Env蛋白以及免疫复合物进行的2-D结构分析进行3-D结构分析来扩展单个分子和复合物的反应点映射研究。主要方法是通过负染色和冷冻电子显微镜（EM）层析成像分析完整的病毒体，以及通过二-D晶体阵列的电子衍射对纯化的重组ENV和免疫复合物的分析。