• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Molecular Biology >Evolution under Drug Pressure Remodels the Folding Free-Energy Landscape of Mature HIV-1 Protease
【24h】

Evolution under Drug Pressure Remodels the Folding Free-Energy Landscape of Mature HIV-1 Protease

机译：药物压力下的进化重塑了成熟HIV-1蛋白酶的折叠自由能态。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Using high-pressure NMR spectroscopy and differential scanning calorimetry, we investigate the folding landscape of the mature HIV-1 protease homodimer. The cooperativity of unfolding was measured in the absence or presence of a symmetric active site inhibitor for the optimized wild type protease (PR), its inactive variant PRD25N, and an extremely multidrug-resistant mutant, PR20. The individual fit of the pressure denaturation profiles gives rise to first order, Delta G(NMR), and second order, Delta V-NMR (the derivative of Delta G(NMR) with pressure); apparent thermodynamic parameters for each amide proton considered. Heterogeneity in the apparent Delta V-NMR values reflects departure from an ideal cooperative unfolding transition. The narrow to broad distribution of Delta V-NMR spanning the extremes from inhibitor-free PR20(D25N) to PR-DMP323 complex, and distinctively for PRD25N-DMP323 complex, indicated large variations in folding cooperativity. Consistent with this data, the shape of thermal unfolding transitions varies from asymmetric for PR to nearly symmetric for PR20, as dimer-inhibitor ternary complexes. Lack of structural cooperativity was observed between regions located close to the active site, including the hinge and tip of the glycine-rich flaps, and the rest of the protein. These results strongly suggest that inhibitor binding drastically decreases the cooperativity of unfolding by trapping the closed flap conformation in a deep energy minimum. To evade this conformational trap, PR20 evolves exhibiting a smoother folding landscape with nearly an ideal two-state (cooperative) unfolding transition. This study highlights the malleability of retroviral protease folding pathways by illustrating how the selection of mutations under drug pressure remodels the free-energy landscape as a primary mechanism. Published by Elsevier Ltd.
机译：使用高压NMR光谱和差示扫描量热法，我们研究了成熟的HIV-1蛋白酶同源二聚体的折叠态。在不存在或存在针对优化的野生型蛋白酶（PR）的对称活性位点抑制剂，其无活性变异体PRD25N和极耐多药性的突变体PR20的情况下，测量了展开的协同性。压力变性曲线的个体拟合产生一阶Delta G（NMR）和二阶Delta V-NMR（Delta G（NMR）随压力的导数）;每个酰胺质子的表观热力学参数。表观δV-NMR值的异质性反映了偏离理想的协同展开过渡。从无抑制剂的PR20（D25N）到PR-DMP323络合物的极值，从极值到窄幅的Delta V-NMR分布，以及PRD25N-DMP323络合物的不同，表明折叠合作性有很大差异。与此数据一致，热展开转变的形状从PR的不对称变化到PR20的近对称变化，为二聚体抑制剂三元复合物。在靠近活性位点的区域之间，包括富含甘氨酸的皮瓣的铰链和尖端，以及其余的蛋白质之间，观察到缺乏结构上的协同作用。这些结果有力地表明，抑制剂的结合通过将闭合的皮瓣构象捕获在最小的能量最小值中而大大降低了展开的协同作用。为了逃避这种构象陷阱，PR20不断发展，展现出一种更为平滑的折叠环境，并具有理想的两态（合作）展开过渡。这项研究通过说明在药物压力下选择突变如何重塑自由能格局作为主要机制，突出了逆转录病毒蛋白酶折叠途径的可塑性。由Elsevier Ltd.发布

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号