State Key Laboratory of Microbial Metabolism (Shanghai Jiao Tong University), Luc Montagnier Biomedical Research Institute, and College of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai Minhang District, China.
J Chem Inf Model. 2011 Jun 27;51(6):1336-46. doi: 10.1021/ci200112b. Epub 2011 May 20.
The human cytochrome P450 1A2 is an important drug metabolizing and procarcinogen activating enzyme. An experimental study found that a peripheral mutation, F186L, at ∼26 Å away from the enzyme's active site, caused a significant reduction in the enzymatic activity of 1A2 deethylation reactions. In this paper, we explored the effects of this mutation by carrying out molecular dynamics simulations and structural analyses. We found that the long-range effects of the F186L mutation were through a change in protein flexibility and a collective protein motion that caused the main substrate access channel to be mostly closed in the mutant. Our work is the first that combined both access channel analysis and protein motion analysis to elucidate mechanisms of mutation-induced allostery in a CYP protein. Such structural modeling and analysis approaches may be applied to other CYP proteins and other enzymes with buried active sites and may help guide protein engineering and drug design.
人细胞色素 P450 1A2 是一种重要的药物代谢和前致癌物激活酶。一项实验研究发现，距酶活性位点约 26Å 的外周突变 F186L 导致 1A2 脱乙基反应的酶活性显著降低。在本文中，我们通过进行分子动力学模拟和结构分析来探索该突变的影响。我们发现，F186L 突变的远程效应是通过改变蛋白质的灵活性和集体蛋白质运动，导致突变体中的主要底物进入通道大部分关闭。我们的工作首次将进入通道分析和蛋白质运动分析相结合，阐明了 CYP 蛋白中突变诱导变构的机制。这种结构建模和分析方法可应用于其他 CYP 蛋白和其他具有埋藏活性位点的酶，并可能有助于指导蛋白质工程和药物设计。
