Mutational effects on the catalytic mechanism of cellobiohydrolase I from Trichoderma reesei.

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	Mutational effects on the catalytic mechanism of cellobiohydrolase I from Trichoderma reesei.
	Yan, S.; Li, T.; & Yao, L
	2011
发表期刊	J Phys Chem B
卷号	115 期号:17 页码:4982-4989
摘要	ABSTRACT: QM/MD simulations are performed to study mutational effects on the glycosylation step of the oligosaccharide hydrolysis catalyzed by Trichoderma reesei cellobiohydrolase I. The potential of mean force along the reaction pathway is determined by the umbrella sampling method. A detailed mechanism is developed to illustrate the decrease in activity of the mutants. Our calculations demonstrate that (1) the E212Q mutation increases the overall activation barrier by ∼4.0 kcal/mol, while the D214N mutation causes∼0.4 kcal/mol increase of the barrier, and (2) there is only one transition state identified in the wild type (WT) and D214N mutant, while two transition states exist in the E212Q mutant for the glycosylation process. The results explain the experimental observation that the E212Q mutant loses most of its hydrolysis capability, while the D214N mutant only reduces it slightly compared to the WT. Further analysis suggests that the proton transfer from Glu217 to O4 and the glycosidic bond cleavage between subsitest1 and_1 are concerted, facilitating the subsequent nucleophilic attack of Glu212 on C1 0 in subsite _1. Our QM/MD study illustrates the importance of the prearrangement of the active site and provides atomic details of the enzymatic catalytic mechanism.
学科领域	仿真与模拟
收录类别	SCI
语种	英语
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/1732
专题	蛋白质设计研究组
推荐引用方式 GB/T 7714	Yan, S.,Li, T.,& Yao, L. Mutational effects on the catalytic mechanism of cellobiohydrolase I from Trichoderma reesei.[J]. J Phys Chem B,2011,115(17):4982-4989.
APA	Yan, S.,Li, T.,&& Yao, L.(2011).Mutational effects on the catalytic mechanism of cellobiohydrolase I from Trichoderma reesei..J Phys Chem B,115(17),4982-4989.
MLA	Yan, S.,et al."Mutational effects on the catalytic mechanism of cellobiohydrolase I from Trichoderma reesei.".J Phys Chem B 115.17(2011):4982-4989.

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