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	Dynamic redox environment-intensified disulfide bond shuffling for protein refolding in vitro: molecular simulation and experimental validation
	Diannan Lu ; Zheng Liu
	2010-10-12 ; 2010-10-12
关键词	Experimental/ biochemistry enzymes hydrophobicity molecular biophysics molecular configurations oxidation proteins reduction (chemical)/ protein refolding molecular simulation disulfide bond formation molecular bond shuffling protein conformational stability two-step folding mechanism hydrophobic collapse molecular structural rearrangement hydrophobic core dynamic redox environment oxidized glutathione hen-egg lysozyme urea concentration GSSG-GSH ratio/ A8715B Biomolecular structure, configuration, conformation, and active sites A8715D Physical chemistry of biomolecular solutions condensed states A3620C Macromolecular conformation (statistics and dynamics) A3620H Macromolecular configuration (bonds, dimensions) A3620E Macromolecular constitution (chains and sequences) A8715H Biomolecular dynamics, molecular probes, molecular pattern recognition
中文摘要	One challenge in protein refolding is to dissociate the non-native disulfide bonds and promote the formation of native ones. In this study, we present a coarse-grained off-lattice model protein containing disulfide bonds and simulate disulfide bond shuffling during the folding of this model protein. Introduction of disulfide bonds in the model protein led to enhanced conformational stability but reduced foldability in comparison to counterpart protein without disulfide bonds. The folding trajectory suggested that the model protein retained the two-step folding mechanism in terms of hydrophobic collapse and structural rearrangement. The disulfide bonds located in the hydrophobic core were formed before the collapsing step, while the bonds located on the protein surface were formed during the rearrangement step. While a reductive environment at the initial stage of folding favored the formation of native disulfide bonds in the hydrophobic core, an oxidative environment at a later stage of folding was required for the formation of disulfide bonds at protein surface. Applying a dynamic redox environment that is, one that changes from reductive to oxidative, intensified disulfide bond shuffling and thus resulted in improved recovery of the native conformation. The above-mentioned simulation was experimentally validated by refolding hen-egg lysozyme at different urea concentrations and oxidized glutathione/reduced glutathione (GSSG/GSH) ratios, and an optimal redox environment, in terms of the GSSG to GSH ratio, was identified. The implementation of a dynamic redox environment by tuning the GSSG/GSH ratio further improved the refolding yield of lysozyme, as predicted by molecular simulation.
语种	英语
出版者	American Chemical Society ; USA
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/79505]
专题	清华大学
推荐引用方式 GB/T 7714	Diannan Lu,Zheng Liu. Dynamic redox environment-intensified disulfide bond shuffling for protein refolding in vitro: molecular simulation and experimental validation[J],2010, 2010.
APA	Diannan Lu,&Zheng Liu.(2010).Dynamic redox environment-intensified disulfide bond shuffling for protein refolding in vitro: molecular simulation and experimental validation..
MLA	Diannan Lu,et al."Dynamic redox environment-intensified disulfide bond shuffling for protein refolding in vitro: molecular simulation and experimental validation".(2010).

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