title: Conformational changes and inactivation of rabbit muscle creatine kinase in dimethyl sulfoxide solutions.
abstract: The effects of dimethyl sulfoxide (DMSO) on creatine kinase (CK) conformation and enzymatic activity were studied by measuring activity changes, aggregation, and fluorescence spectra. The results showed that at low concentrations (< 65% v/v), DMSO had little effect on CK activity and structure. However, higher concentrations of DMSO led to CK inactivation, partial unfolding, and exposure of hydrophobic surfaces and thiol groups. DMSO caused aggregation during CK denaturation. A 75% DMSO concentration induced the most significant aggregation of CK. The CK inactivation and unfolding kinetics were single phase. The unfolding of CK was an irreversible process in the DMSO solutions. The results suggest that to a certain extent, an enzyme can maintain catalytic activity and conformation in water-organic mixture environments. Higher concentrations of DMSO affected the enzyme structure but not its active site. Inactivation occurred along with noticeable conformational change during CK denaturation. The inactivation and unfolding of CK in DMSO solutions differed from other denaturants such as guanidine, urea, and sodium dodecyl sulfate. The exposure of hydrophobic surfaces was a primary reason for the protein aggregation.
authors.affiliation: Department of Biological Science and Biotechnology, Tsinghua University, Beijing, PR China.
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Ou
authors.forename: Ri-Sheng
authors.initials: RS
authors.lastname: Wang
authors.forename: Hai-Meng
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000818:Animals; D003402:Creatine Kinase / Q000737:chemistry*; D004121:Dimethyl Sulfoxide / Q000737:chemistry*; D004228:Dithionitrobenzoic Acid / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D007700:Kinetics; D009132:Muscles / Q000737:chemistry / Q000201:enzymology*; D011487:Protein Conformation; D011489:Protein Denaturation; D020673:Protein Renaturation; D011817:Rabbits
chemical_list: D004228:Dithionitrobenzoic Acid; D003402:Creatine Kinase; D004121:Dimethyl Sulfoxide