title: High salt concentrations induce dissociation of dimeric rabbit muscle creatine kinase. Physico-chemical characterization of the monomeric species.
abstract: Incubation of dimeric MM-creatine kinase (MM-CK) with high NaCl or LiCl concentrations results in dissociation of the subunits and complete enzyme inactivation. In NaCl, this process, which depends on protein concentration, may be described according to a two-state model where the dimer can be reversibly converted into compact folded monomers (D <--> 2M). At LiCl concentrations higher than 2-2.5 M, MM-CK is recovered in two monomeric states: an inactive compact species (M) and a more expanded form (EF), which represents 15-20% of the population. Thus, in LiCl, a three-state model (D <--> 2M --> 2EF) more adequately accounts for our experimental results. The monomeric species (M) obtained in NaCl and LiCl exhibits some properties of the molten globule state described in guanidine hydrochloride. Indeed, this form is compact and devoid of any enzymatic activity; it maintains a high degree of secondary structure and binds 8-anilino-1-naphthalenesulfonate. The formation of this intermediate induces the exposure of a second tryptophan (among the four present) which is located at the monomer-monomer interface in the native structure. In LiCl, the monomeric species (M) is irreversibly converted into a less compact form (EF) which seems to have lost a large part of its secondary structure.
authors.affiliation: UPRESA 5013 CNRS-Lyon I, Biomembranes et Enzymes Associés, Villeurbanne, France.
authors.forename: F
authors.initials: F
authors.lastname: Couthon
authors.forename: E
authors.initials: E
authors.lastname: Clottes
authors.forename: C
authors.initials: C
authors.lastname: Vial
mesh_terms: D000818:Animals; D001665:Binding Sites; D003402:Creatine Kinase / Q000737:chemistry*; D019281:Dimerization; D018021:Lithium Chloride; D018482:Muscle, Skeletal / Q000201:enzymology*; D011487:Protein Conformation; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D011817:Rabbits; D012462:Saline Solution, Hypertonic
chemical_list: D012462:Saline Solution, Hypertonic; D003402:Creatine Kinase; D018021:Lithium Chloride