title: Unfolding, conformational change of active sites and inactivation of creatine kinase in SDS solutions.
abstract: It has been reported that inactivation occurs before noticeable conformational change can be detected during denaturation of creatine kinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2) and other enzymes by guanidinium chloride or urea. Therefore, Tsou suggested that enzyme active sites may display more conformational flexibility than the enzyme molecules as a whole [Tsou (1986) Trends Biochem. Sci. 11, 427-429; Tsou (1993) Science 262, 380-381]. In this study, the conformational change of the active site, the unfolding of the whole molecule and the inactivation of creatine kinase in solutions of different concentrations of SDS are compared. The results show that, at low SDS concentrations, the conformational change of the active site and inactivation of the enzyme occur to nearly the same extent. However, both of these changes occur at much lower concentrations of SDS than required to significantly unfold the enzyme molecule. The rates of conformational changes of enzyme active sites are markedly faster than those of inactivation. However, at the same SDS concentration, both the inactivation rate and the rate of the active site conformational change are much faster than that of the unfolding of the enzyme molecule as a whole. The above results provide direct evidence of the flexibility of the active site of creatine kinase.
authors.affiliation: Department of Biological Science and Biotechnology, Tsinghua University, Beijing, People's Republic of China.
authors.forename: Z F
authors.initials: ZF
authors.lastname: Wang
authors.forename: M Q
authors.initials: MQ
authors.lastname: Huang
authors.forename: X M
authors.initials: XM
authors.lastname: Zou
authors.forename: H M
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001665:Binding Sites; D003402:Creatine Kinase / Q000737:chemistry*; D009132:Muscles / Q000201:enzymology; D011487:Protein Conformation; D017510:Protein Folding; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate; D012996:Solutions
chemical_list: D012996:Solutions; D012967:Sodium Dodecyl Sulfate; D003402:Creatine Kinase