title: Ascertaining the number of essential thiol groups for the folding of creatine kinase.
abstract: Although the unfolding and refolding of proteins have been extensively studied in the 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 (Sci. Sin. 1962, 11, 1535-1558) 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 cystein 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 cystein residue modified in each subunit of creatine kinase, only three are essential for its folding.
authors.affiliation: Department of Biological Science and Biotechnology, Tsinghua University, Beijing, People's Republic of China.
authors.forename: H R
authors.initials: HR
authors.lastname: Wang
authors.forename: J H
authors.initials: JH
authors.lastname: Bai
authors.forename: S Y
authors.initials: SY
authors.lastname: Zheng
authors.forename: Z X
authors.initials: ZX
authors.lastname: Wang
authors.forename: H M
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000818:Animals; D002730:Chloromercurinitrophenols / Q000737:chemistry; D003402:Creatine Kinase / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000737:chemistry
chemical_list: D002730:Chloromercurinitrophenols; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; C026149:2-chloromercuri-4-nitrophenol; D003402:Creatine Kinase