Assembly of HIV intasomes

HIV整合体的组装

• 批准号: 9203230
• 负责人: DUANE P GRANDGENETT
• 金额: $ 18.94万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-15 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9203230
• 关键词: Active Sites; Binding; Biochemical; Catalysis; Catalytic Domain; Complex; Crystal Formation; Crystallization; DNA; DNA Sequence; DNA Structure; Development; Distal; Drug resistance; Future; Genome; Goals; HIV; HIV Integrase; HIV-1; Harvest; In Vitro; Individual; Integrase; Integrase Inhibitors; Investigation; Length; Methods; Modeling; Pathway interactions; Prophylactic treatment; Proteins; Recombinants; Resolution; Retroviridae; Robotics; Roentgen Rays; Role; Rous sarcoma virus; Site; Solubility; Spumavirus; Structure; Synapses; System; Viral; abstracting; base; biophysical analysis; dimer; inhibitor/antagonist; insight; light scattering; novel; protein function; prototype; screening; success; three dimensional structure; viral DNA

Project Abstract: The X-ray structures of the 4-domain prototype foamy virus (PFV) integrase (IN)(1,2) and very recently the 3-domain Rous sarcoma virus (RSV) IN (3) in complex with viral DNA and/or viral-target DNA substrates have been determined. Yet, the crystal structure of the medically important 3- domain HIV IN by itself or in complex with DNA is still elusive. We propose to provide the means and methods using monomeric wild type (wt) HIV IN or modified IN constructs to accomplish this very difficult but highly significant project. The first necessary step to achieve this important goal is harvesting the ability of HIV IN to assemble these IN-DNA complexes at high protein concentrations in the ~100 to 150 µM range for crystal screening and other in-solution structural investigations. Our recent success in defining new assembly pathways for stabilizing 3-domain IN-DNA complexes and the use of newly enhanced solubility methods for recombinant HIV IN will advance our investigative studies.

项目摘要： 泡沫病毒4结构域原型整合酶(IN)(1，2)和Very的X射线结构 最近发现的3区劳斯肉瘤病毒(RSV)IN(3)与病毒DNA和/或病毒靶标形成复合体 DNA底物已确定。然而，医学上重要的3种化合物的晶体结构- HIVIN本身或与DNA的复合体仍然难以捉摸。我们建议提供所需资源和 方法使用单体野生型(Wt)HIV IN或修饰的IN构建来实现这一点 难度很大但意义重大的项目。实现这一重要目标的第一个必要步骤是 获得HIV IN在高蛋白浓度下组装这些IN-DNA复合体的能力 在~100至150微米范围内用于晶体筛选和其他溶液结构研究。我们的 最近成功地定义了稳定3-结构域IN-DNA复合体的新组装途径和 使用新增强的溶解度方法检测重组HIV-IN将推动我们的研究 学习。