SMALL ANGLE X-RAY SCATTERING STUDIES OF HIV NUCLEOPROTEIN

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 虽然蛋白质数据库中有HIV整合酶的几个片段，但完整的全长整合酶以其催化相关的寡聚态和整合酶-DNA复合体的实验结构一直没有得到结晶学分析。这项研究的目标是开发这些大分子复合体的实验衍生模型，这对于促进我们对HIV整合及其结构基础的理解至关重要。目前的艾滋病大流行使设计有效的药物治疗成为最重要的目标。虽然一些有希望的整合酶抑制剂是从最近的临床试验中出现的，但大多数可用于临床的抑制剂只针对其他病毒编码的蛋白质。由于病毒c DNA整合到宿主基因组中对逆转录病毒的生命周期至关重要，而且目前还没有已知的在序列或功能上与整合酶非常相似的细胞酶，整合酶抑制剂有可能是相对无毒和非常有效的。现在清楚的是，与溶液中的游离整合酶相比，正确组装的整合酶复合体对抑制剂具有不同的和更具区分性的反应。因此，一个详细的复合体结构模型对于基于结构的药物设计是至关重要的。HIV整合酶、整合酶结合蛋白晶状体上皮源性生长因子(LEDGF)和DNA底物形成的复合体的生物物理特性已经通过分析超速离心和凝胶过滤完成。在国际象棋的光束线G1进行的SAXS试点实验已经在有限的样品浓度范围内进行。

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.