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Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase
He, Hua-Wei ; Li, He ; Zhao, Tong-Jin ; Ma, Yong ; Shi, Feng ; Zhou, Hai-Meng
2010-05-11 ; 2010-05-11
关键词creatine kinase inactivation kinetics chemical modification cysteine STATE ANALOG COMPLEX ACTIVE-SITE SUBSTRATE REACTION THIOL-GROUPS KINETICS INACTIVATION PK(A) RESIDUE VALUES DOMAIN Biochemistry & Molecular Biology
中文摘要The polar microenvironment around the reactive Cys283 of rabbit muscle creatine kinase was explored using kinetic analysis of substrates reaction in the presence of modifiers. In the present study, three specific sulphydryl reagents, 5,5'-dithiobis(2-nitrobenzoic acid), 6,6'-dithiodinicotinic acid and 2,2'-dithiodipyridine, were applied as modifiers to react with Cys283 of creatine kinase. The inactivation kinetics of creatine kinase by the modifiers was analyzed. The microscopic rate constants for reactions of the modifiers with free enzyme and enzyme-substrate complexes were also determined. The results suggested that the inactivation rate of creatine kinase by 5,5'-dithiobis(2-nitrobenzoic acid) was the fastest, followed by 6,6'-dithiodinicotinic acid and then 2,2'-dithiodipyridine. Interestingly, 5,5'-dithiobis(2-nitrobenzoic acid) and 6,6'-dithiodinicotinic acid functioned as non-complexing modifiers, while 2,2'-dithiodipyridine did a complexing modifier. The results here indicated that the electrophilic group was predominant around Cys283, and that the presence of substrates seemed to have different effects on the inactivation reactions of creatine kinase by the three modifiers. Furthermore, the findings in this study may provide a novel explanation for the low pK(a) value of Cys283. (C) 2007 Elsevier B.V. All rights reserved.
语种英语 ; 英语
出版者ELSEVIER SCIENCE BV ; AMSTERDAM ; PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
内容类型期刊论文
源URL[http://hdl.handle.net/123456789/26474]  
专题清华大学
推荐引用方式
GB/T 7714
He, Hua-Wei,Li, He,Zhao, Tong-Jin,et al. Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase[J],2010, 2010.
APA He, Hua-Wei,Li, He,Zhao, Tong-Jin,Ma, Yong,Shi, Feng,&Zhou, Hai-Meng.(2010).Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase..
MLA He, Hua-Wei,et al."Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase".(2010).
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