Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase

Hua Wei He, Jie Li, Tong Jin Zhao, Yong Ma, Feng Shi, Hai Meng Zhou

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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 pKa value of Cys283.

Original languageEnglish (US)
Pages (from-to)361-368
Number of pages8
JournalInternational Journal of Biological Macromolecules
Volume41
Issue number4
DOIs
StatePublished - Oct 1 2007

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MM Form Creatine Kinase
Nitrobenzoates
Creatine Kinase
Cysteine
Rabbits
Acids
Substrates
Kinetics
Enzymes
Rate constants

Keywords

  • Chemical modification
  • Creatine kinase
  • Cysteine
  • Inactivation
  • Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase. / He, Hua Wei; Li, Jie; Zhao, Tong Jin; Ma, Yong; Shi, Feng; Zhou, Hai Meng.

In: International Journal of Biological Macromolecules, Vol. 41, No. 4, 01.10.2007, p. 361-368.

Research output: Contribution to journalArticle

He, Hua Wei ; Li, Jie ; Zhao, Tong Jin ; Ma, Yong ; Shi, Feng ; Zhou, Hai Meng. / Exploration of the polar microenvironment around the reactive cysteine in rabbit muscle creatine kinase. In: International Journal of Biological Macromolecules. 2007 ; Vol. 41, No. 4. pp. 361-368.
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