Computational redesign of human butyrylcholinesterase for anticocaine medication
Computational redesign of human butyrylcholinesterase for anticocaine medication
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DOI：
10.1073/pnas.0507332102
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发表时间：
2005-11-15
影响因子：
11.1
通讯作者：
Zhan, CG
中科院分区：
文献类型：
--
作者：
Pan, YM;Gao, DQ;Zhan, CG
用分子动力学方法模拟了人丁酰胆碱酯酶（BChE）及其突变体催化可卡因水解的第一步化学反应（TS 1）的过渡态。模拟结果表明，A199 S/S287 G/A328 W/Y332 G BChE催化（-）-可卡因水解的TS 1结构中，（-）-可卡因苯甲酰酯的羰基氧与BChE的氧阴离子空穴之间的整体氢键应明显强于WT BChE和其他模拟BChE突变体催化（-）-可卡因水解的TS 1结构中的氢键.因此，过渡态模拟预测BChE的A199 S/S287 G/A328 W/Y332 G突变体对于反应过程应该具有显著较低的能垒，并且因此对于（-）-可卡因水解具有显著较高的催化效率。理论预测已被湿实验测试所证实，表明A199 S/S287 G/A328 W/Y332 G BChE对（-）-可卡因的催化效率提高了约（456 +/-41）倍。这是一个独特的研究，设计一个基于过渡态模拟的酶突变体。所设计的BChE突变体对可卡因的催化效率最高的所有报告的BChE突变体，表明基于过渡态模拟的独特的设计方法是有希望的合理的酶的重新设计和药物发现。
Molecular dynamics was used to simulate the transition state for the first chemical reaction step (TS1) of cocaine hydrolysis catalyzed by human butyrylcholinesterase (BChE) and its mutants. The simulated results demonstrate that the overall hydrogen bonding between the carbonyl oxygen of (-)-cocaine benzoyl ester and the oxyanion hole of BChE in the TS1 structure for (-)-cocaine hydrolysis catalyzed by A199S/S287G/A328W/Y332G BChE should be significantly stronger than that in the TS1 structure for (-)-cocaine hydrolysis catalyzed by the WT BChE and other simulated BChE mutants. Thus, the transition-state simulations predict that A199S/ S287G/A328W/Y332G mutant of BChE should have a significantly lower energy barrier for the reaction process and, therefore, a significantly higher catalytic efficiency for (-)-cocaine hydrolysis. The theoretical prediction has been confirmed by wet experimental tests showing an approximate to (456 +/- 41)-fold improved catalytic efficiency of A199S/S287G/A328W/Y332G BChE against (-)-cocaine. This is a unique study to design an enzyme mutant based on transition-state simulation. The designed BChE mutant has the highest catalytic efficiency against cocaine of all of the reported BChE mutants, demonstrating that the unique design approach based on transition-state simulation is promising for rational enzyme redesign and drug discovery.