STRUCTURE BASED THERMODYNAMIC STUDIES OF HIV1 PROTEASE

描述：（改编自申请人的摘要）：本补助金申请 重点是阐明临床耐药的分子基础， HIV-1 PR抑制剂。 HIV-1 PR的耐药突变体显示出较低的亲和力， 抑制剂，但仍然能够处理病毒多蛋白底物， 病毒复制的过程 抗性的氨基酸序列 从临床研究中已知突变体。 为了阐明本 对HIV-1蛋白酶抑制剂耐药的分子基础，完整的 底物和抑制剂与HIV-1 PR的结合能（wt; 抗性突变体）将通过高灵敏度反应来测量 量热技术和标准光谱分析。 基于结构 热力学分析将使用一种新的方法开发， 申请人的实验室。 这些信息将用于开发一个 底物和抑制剂结合能的结构图。 的 结构图将确定每个组的能量贡献， 蛋白酶、底物或抑制剂分子对整体结合 亲和力 该地图还将确定焓和熵的贡献 结合能量学，并将熵贡献分解为 构象和溶剂相关组分。 类型的差异和 相互作用的强度将被评估，并用于确定为什么一些 特定的突变影响底物的结合亲和力， 以不同的方式抑制。

DESCRIPTION: (Adapted from applicant's Abstract): This grant application is focused on elucidating the molecular basis of clinical resistance to HIV-1 PR inhibitors. Resistant mutants of HIV-1 PR show lower affinity for inhibitors, yet are still able to process viral polyprotein substrates in the course of viral replication. The amino acid sequences of resistant mutants are known from clinical studies. In order to elucidate the molecular basis of resistance to HIV-1 protease inhibitors, the complete binding energetics of substrate and inhibitors to the HIV-1 PR (wt; resistance mutants) will be measured by high sensitivity reaction calorimetric techniques and standard spectroscopic assays. Structure-based thermodynamic analysis will be performed using a novel approach developed in the applicant's laboratory. This information will be used to develop a structural map of the binding energetics of substrates and inhibitors. The structural map will identify the energetic contributions from each group in the protease, substrate or inhibitor molecules to the overall binding affinity. This map will also identify enthalpic and entropic contributions to the binding energetics, and dissect the entropy contributions into conformation and solvent-related components. Differences in the type and strength of interactions will be evaluated and used to identify why some specific mutations affect the binding affinity of the substrate and inhibitor in different ways.