STRUCTURAL BIOLOGY OF HIV ENVELOPE INTERACTIONS

HIV 包膜相互作用的结构生物学

• 批准号: 6336263
• 负责人: STEPHEN COPLAN HARRISON
• 金额: $ 17.2万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6336263
• 关键词: CD4 molecule; HIV envelope protein gp120; HIV envelope protein gp160; HIV envelope protein gp41; X ray crystallography; antiAIDS agent; chemical binding; circular dichroism; conformation; crystallization; fluorescence polarization; human immunodeficiency virus 1; inhibitor /antagonist; ligands; monoclonal antibody; peptide analog; peptide library; receptor binding; structural biology; surface plasmon resonance; virus infection mechanism; virus receptors

There are two sets of aims in this parts of the program project. The first set concerns the CD4/gp120 interaction and attendant conformation changes in gp120. The second set concerns crystallographic analyses of gp120/CD4 complexes and forms of truncated gp160 trimers, prepared as described in part 1. (1) Phage display has been used to identify a group of peptides that inhibit CD4 binding with gp120. The interactions of these peptides with gp120 will be characterized using fluorescence polarization, surface plasmon resonance, circular dischrosm, effects on mAb binding, and related approaches, in order to identify the key determinants of binding/inhibition, to discover tighter-binding peptides, and to determine whether peptide binding affects V3 loop conformation. Crystals of gp120/peptide complexes will be sought. A screen for peptide inhibitors will be undertaken, using bead-based chemical libraries biased by information from the peptide studies. (2) X-ray structural analysis of HIV envelope components will be undertaken, using crystals obtained through the efforts in part 1. Complexes that will be investigated included gp120/CD4 Fab (using gp120 with homogenous, short oligosaccharides expressed in Lec 3.2.8.1 cells) and trimeric ectodomain of gp160. Complexes of small-molecule ligands with gp41, identified by NMR methods in project 3, will also be examined crystallographically.

该计划项目的这一部分有两套目标。第一个 集涉及 CD4/gp120 相互作用和伴随的构象变化 在GP120中。第二组涉及 gp120/CD4 的晶体学分析 截短的 gp160 三聚体的复合物和形式，按照中所述制备 第 1 部分. (1) 噬菌体展示已用于鉴定一组肽 抑制 CD4 与 gp120 的结合。这些肽的相互作用 gp120 将使用荧光偏振、表面 等离振子共振、圆二色性、对 mAb 结合的影响以及相关 方法，以确定关键决定因素 结合/抑制，发现结合更紧密的肽，并确定 肽结合是否影响 V3 环构象。的晶体 将寻找 gp120/肽复合物。肽抑制剂的筛选 将使用基于珠子的化学库进行，偏向于 来自肽研究的信息。 (2) HIV的X射线结构分析 将使用通过获得的晶体来进行包络组件 第 1 部分中的工作。将要研究的复合物包括 gp120/CD4 Fab（使用 gp120 和均质短寡糖 在 Lec 3.2.8.1 细胞中表达）和 gp160 的三聚体胞外域。 通过 NMR 方法鉴定小分子配体与 gp41 的复合物 在项目 3 中，还将进行晶体学检查。