Ascertaining the number of essential thiol groups for the folding of creatine kinase

Hong Rui Wang, Ji Hong Bai, Siyang Zheng, Zhi Xin Wang, Hai Meng Zhou

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Although the unfolding and refolding of proteins have been extensively studied in tile literature, relatively few attempts have been made to see how many residues of the total residues of a certain amino acid in an enzyme can be modified without seriously affecting its folding. Based on a statistical analysis of the quantitative relationship between the extent of modification of protein functional groups and the decrease in their biological activity, a method proposed by Tsou is widely used to determine the number of residues essential for the catalytic activity of modified proteins. In the present paper, Tsou's method is applied to determine the number of cysteine residues essential for the folding of creatine kinase. The thiol groups of the cysteine residues in fully unfolded creatine kinase were modified by 2-chloromercuri-4-nitrophenol (MNP). The relationship between the number of MNP-groups introduced and the recovery of activity after refolding was determined. Quantitative treatment of the data by Tsou's plot shows that among the cysteine residues modified in each subunit of creatine kinase, only three are essential for its folding.

Original languageEnglish (US)
Pages (from-to)174-180
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume221
Issue number1
DOIs
StatePublished - Apr 5 1996

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Creatine Kinase
Sulfhydryl Compounds
Cysteine
Protein Refolding
Protein Unfolding
Proteins
Tile
Bioactivity
Functional groups
Catalyst activity
Statistical methods
Amino Acids
Recovery
Enzymes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wang, Hong Rui ; Bai, Ji Hong ; Zheng, Siyang ; Wang, Zhi Xin ; Zhou, Hai Meng. / Ascertaining the number of essential thiol groups for the folding of creatine kinase. In: Biochemical and Biophysical Research Communications. 1996 ; Vol. 221, No. 1. pp. 174-180.
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Ascertaining the number of essential thiol groups for the folding of creatine kinase. / Wang, Hong Rui; Bai, Ji Hong; Zheng, Siyang; Wang, Zhi Xin; Zhou, Hai Meng.

In: Biochemical and Biophysical Research Communications, Vol. 221, No. 1, 05.04.1996, p. 174-180.

Research output: Contribution to journalArticle

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