SMALL ANGLE X-RAY SCATTERING STUDIES OF HIV NUCLEOPROTEIN

HIV 核蛋白的小角 X 射线散射研究

• 批准号: 8170111
• 负责人: KUSHOL GUPTA
• 金额: $ 0.1万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170111
• 关键词: Acquired Immunodeficiency Syndrome; Binding Proteins; Clinical; Clinical Trials; Complementary DNA; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA; Databases; Drug Design; Enzymes; Epithelial; Funding; Gel Chromatography; Genome; Goals; Grant; Growth Factor; HIV; HIV Integrase; Institution; Integrase; Integrase Inhibitors; Length; Life Cycle Stages; Macromolecular Complexes; Modeling; Nucleoproteins; Pharmacological Treatment; Proteins; Research; Research Personnel; Resources; Sampling; Solutions; Source; Structural Models; Structure; United States National Institutes of Health; Viral; analytical ultracentrifugation; base; beamline; design; inhibitor/antagonist; lens; pandemic disease; research study; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. While several segments of HIV integrase are available in the Protein Data Bank, experimental structures of intact full-length integrase in its catalytically relevant oligomeric states and integrase-DNA complex have eluded crystallographic analysis. The goal of this study is to develop experimentally-derived models of these macromolecular complexes, which is vital to advancing our understanding of HIV integration and its structural basis. The ongoing AIDS pandemic has made the design of effective pharmacological treatments a goal of paramount importance. While some promising integrase inhibitors are emerging from late clinical trials, most available inhibitors in clinical use only target other viral-encoded proteins. Because integration of viral cDNA into the host genome is critical to the retroviral life cycle and since there are no known cellular enzymes that closely resemble integrase in sequence or function, inhibitors of integrase have the potential to be relatively nontoxic and very effective. It is now clear that, in comparison to free integrase in solution, correctly assembled integrase complex has a different and more discriminating response to inhibitors. Hence, a detailed structural model of the complex is essential to structure-based drug design. Biophysical characterization of complexes formed by HIV integrase, the integrase-binding protein Lens Epithelial-Derived Growth Factor (LEDGF), and DNA substrate, using analytical ultracentrifugation and gel filtration, has already been completed. Pilot SAXS experiments at Beamline G1 of CHESS have already been performed at a limited range of sample concentrations.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 虽然蛋白质数据库中提供了 HIV 整合酶的几个片段，但催化相关寡聚状态的完整全长整合酶和整合酶-DNA 复合物的实验结构却未能进行晶体学分析。 这项研究的目标是开发这些大分子复合物的实验衍生模型，这对于增进我们对 HIV 整合及其结构基础的理解至关重要。 持续的艾滋病大流行使得设计有效的药物治疗成为最重要的目标。虽然后期临床试验中出现了一些有前景的整合酶抑制剂，但临床使用的大多数可用抑制剂仅针对其他病毒编码蛋白。由于病毒 cDNA 整合到宿主基因组中对于逆转录病毒生命周期至关重要，并且由于没有已知的细胞酶在序列或功能上与整合酶非常相似，因此整合酶抑制剂具有相对无毒且非常有效的潜力。 现在很清楚，与溶液中的游离整合酶相比，正确组装的整合酶复合物对抑制剂具有不同且更具辨别力的反应。 因此，复合物的详细结构模型对于基于结构的药物设计至关重要。 使用分析超速离心和凝胶过滤，已经完成了 HIV 整合酶、整合酶结合蛋白晶状体上皮衍生生长因子 (LEDGF) 和 DNA 底物形成的复合物的生物物理表征。 CHESS 的 Beamline G1 的 SAXS 试点实验已经在有限的样品浓度范围内进行。