STRUCTURE BASED THERMODYNAMIC STUDIES OF HIV1 PROTEASE

HIV1 蛋白酶的基于结构的热力学研究

• 批准号: 2543066
• 负责人: Ernesto Freire
• 金额: $ 27.16万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2543066
• 关键词: antiAIDS agent; aspartic endopeptidases; calorimetry; drug resistance; gene mutation; human immunodeficiency virus 1; protease inhibitor; protein structure; spectrometry; thermodynamics

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.

描述：(改编自申请者摘要)：本次拨款申请 致力于阐明临床耐药的分子基础。 HIV-1 PR抑制剂。HIV-1 PR耐药突变株对病毒的亲和力较低 抑制剂，但仍然能够处理病毒多蛋白底物在 病毒复制的过程。抗病的氨基酸序列 突变是从临床研究中得知的。为了澄清这个问题 HIV-1蛋白酶抑制剂耐药的分子基础 底物和抑制剂与HIV-1PR的结合能量学(wt； 抗性突变体)将通过高灵敏度反应进行测量 量热技术和标准光谱分析。基于结构的 将使用中开发的新方法执行热力学分析 申请人的实验室。这些信息将被用来开发一个 底物和缓蚀剂结合能的结构图。这个 结构图将确定来自每个小组的能量贡献 将蛋白水解酶、底物或抑制物分子全面结合 亲和力。这张地图还将识别出焓和熵的贡献。 到束缚能量学，并将熵的贡献分解为 构象和与溶剂相关的成分。在类型和类型上的差异 互动的强度将被评估并用来确定为什么 特定突变会影响底物的结合亲和力 以不同的方式抑制。