HIV-1 Integrase Structural Biology

HIV-1 整合酶结构生物学

• 批准号: 7579809
• 负责人: Alan N. Engelman
• 金额: $ 40.72万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7579809
• 关键词: Arts; Binding; Binding Sites; Biochemical; Biological Assay; C-terminal; Catalytic Domain; Cells; Chromosomes; Clinical Trials; Complementary DNA; Complex; Crystallization; Crystallography; DNA; Data; Drug Delivery Systems; Drug Design; Enzymes; Equilibrium Centrifugation; Gel Chromatography; Glean; HIV-1; HIV-1 integrase; Hot Spot; Housing; Human; In Vitro; Integrase; Integrase Inhibitors; Isotonic Exercise; Length; Molecular; Molecular Weight; Mutation; N-terminal; NMR Spectroscopy; Pathway interactions; Pattern; Phase; Proteins; Reaction; Retroviridae; Reverse Transcription; Site; Sodium Chloride; Solubility; Source; Structure; Suicide; System; Techniques; Viral; Work; base; design; dimer; enzyme structure; inhibitor/antagonist; killings; lens epithelium-derived growth factor; light scattering; mutant; novel; research study; sedimentation equilibrium; small molecule; structural biology; synchrotron radiation; three dimensional structure; transcriptional coactivator p75

Like all retroviruses, HIV-1 must integrate the cDNA copy made by reverse transcription into a cell chromosome in order to replicate. HIV-1integration is catalyzed by the essential viral enzyme integrase. This makes the integrase an attractive drug target, and anti-integrase compounds are in US clinical trials. Rational drug design benefits from visualizing three-dimensional enzyme structures, and numerous structures of the HIV-1 integrase catalytic core domain have been solved. Although two domain N-terminal/core and core/C-terminal structures are also solved, there is no structure for the intact integrase protein, which in large part can be attributed to poor protein solubility. In cells, integrase is likely to interact with normal human proteins, and the interaction between integrase and the transcriptional co-activator lens epithelium-derived growth factor (LEDGF) forms the basis for this application. We recently solved the three-dimensional structure of the integrase-binding domain (IBD)in LEDGF using NMR spectroscopy and herein present the crystal structure of the IBD bound to the dimeric core domain of HIV-1 integrase. Building from these results, we propose to utilize the LEDGF IBD to form novel LEDGF-IN complexes and to solve the three-dimensional structure of the full-length integrase protein. Our novel structure revealed a pocket at the core domain dimer interface that is occupied by LEDGF hot spot residues upon complex formation. Because integrase mutations that ablate the interaction with LEDGF kill HIV-1, we hypothesize that compounds that bind to this region of the core and preclude LEDGF binding might similarly cripple HIV-1. Previous results revealed that certain integrase inhibitors bind at or near this pocket, but those compounds did not inhibit the LEDGF-integrase interaction. A novel assay system will be designed to select for inhibitors of the LEDGF-integrase interaction. The results of these experiments will significantly aid the design of inhibitors of HIV-1 integrase function, as the three-dimensional structure of the intact enzyme will be elucidated and an assay that could select for novel inhibitors of HIV-1 replication will be developed.

像所有逆转录病毒一样，HIV-1必须将逆转录产生的cDNA拷贝整合到细胞中 染色体，以便复制。HIV-1的整合是由病毒的基本酶催化的 整合酶这使得整合酶成为一个有吸引力的药物靶标，并且抗整合酶化合物在 美国临床试验。三维酶的可视化有助于药物设计的合理化 结构，并且已经解决了HIV-1整合酶催化核心结构域的许多结构。 虽然也解决了两个结构域N-末端/核心和核心/C-末端结构，但没有解决。 完整整合酶蛋白质的结构，这在很大程度上可以归因于蛋白质缺乏 溶解度在细胞中，整合酶可能与正常的人类蛋白质相互作用，并且这种相互作用 整合酶与转录共激活因子透镜上皮源性生长因子之间 （LEDGF）是本申请的基础。我们最近解决了 使用NMR光谱分析LEDGF中的整合酶结合结构域（IBD），并在本文中呈现 IBD的晶体结构结合到HIV-1整合酶的二聚体核心结构域。建筑从 根据这些结果，我们建议利用LEDGF IBD形成新的LEDGF-IN复合物， 解决了全长整合酶蛋白的三维结构。我们的新结构 揭示了核心结构域二聚体界面处的口袋，其被LEDGF热点残基占据 复杂的形成。因为整合酶突变消除了与LEDGF的相互作用， HIV-1，我们假设与核心区域结合并阻止LEDGF的化合物 结合可能同样削弱HIV-1。以前的结果表明，某些整合酶抑制剂结合 在该口袋处或附近，但这些化合物不抑制LEDGF-整合酶相互作用。一种新型 将设计测定系统以选择LEDGF-整合酶相互作用的抑制剂。的 这些实验的结果将显著有助于HIV-1整合酶功能抑制剂的设计， 因为完整酶的三维结构将被阐明， 将开发新型HIV-1复制抑制剂。