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	Role of the linker between the N- and C-terminal domains in the stability and folding of rabbit muscle creatine kinase
	He, Hua-Wei ; Feng, Shan ; Pang, Min ; Zhou, Hai-Meng ; Yan, Yong-Bin
	2010-05-11 ; 2010-05-11
关键词	linker multidomain protein muscle creatine kinase protein folding protein aggregation protein stability M-LINE PROTEIN INTERMEDIATE EQUILIBRIUM DENATURATION REACTIVATION AGGREGATION EVOLUTION SEQUENCE BINDING Biochemistry & Molecular Biology Cell Biology
中文摘要	Domain-domain interactions may be very important to the structure and functions of many multidomain proteins. However, little is known about the role of the linker in the folding, stability and function of multidomain proteins. In this research, muscle creatine kinase (CK), a dimeric two-domain protein, was used as a model protein to investigate the role of the linker in CK activity, stability and folding by mutational analysis. Two of the three mutations, L115D and L121D, resulted in a gradual decrease in CK activity and secondary structures, but did not affect CK inactivation induced by heat or guanidine hydrochloride (GdnHCl). The mutations also caused much more serious aggregation during heat- and GdnHCl-induced denaturation and refolding from the GdnHCl-denatured state. More importantly, none of the three mutants could successfully recover their activities by dilution-initiated refolding, and the rate constant of CK refolding was gradually decreased by the mutations. These results suggested that mutations of the hydrophobic residues in the linker might affect the correct positioning of the domains and thus disrupt the efficient recognition and interactions between the two domains. The results herein indicated that in addition to its role in the in vivo functions, the linker also played a crucial role in the stability and folding of CK. (c) 2007 Elsevier Ltd. All rights reserved.
语种	英语 ; 英语
出版者	PERGAMON-ELSEVIER SCIENCE LTD ; OXFORD ; THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/26615]
专题	清华大学
推荐引用方式 GB/T 7714	He, Hua-Wei,Feng, Shan,Pang, Min,et al. Role of the linker between the N- and C-terminal domains in the stability and folding of rabbit muscle creatine kinase[J],2010, 2010.
APA	He, Hua-Wei,Feng, Shan,Pang, Min,Zhou, Hai-Meng,&Yan, Yong-Bin.(2010).Role of the linker between the N- and C-terminal domains in the stability and folding of rabbit muscle creatine kinase..
MLA	He, Hua-Wei,et al."Role of the linker between the N- and C-terminal domains in the stability and folding of rabbit muscle creatine kinase".(2010).

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