Effects of Arginine on Rabbit Muscle Creatine Kinase and Salt-Induced Molten Globule-Like State


The arginine (Arg)-induced unfolding and the salt-induced folding of creatine kinase (CK) have been studied by measuring enzyme activity, fluorescence emission spectra, native polyacrylamide gel electrophoresis and size exclusion chromatography (SEC). The results showed that Arg caused inactivation and unfolding of CK, but there was no aggregation during CK denaturation. The kinetics of CK unfolding followed a one-phase process. At higher concentrations of Arg (>160 mM), the CK dimers were fully dissociated, the alkali characteristic of Arg mainly led to the dissociation of dimers, but not denaturation effect of Arg's guanidine groups on CK. The inactivation of CK occurred before noticeable conformational changes of the whole molecules. KCl induced monomeric and dimeric molten globule-like states of CK denatured by Arg. These results suggest that as a protein denaturant, the effect of Arg on CK differed from that of guanidine and alkali, its denaturation for protein contains the double effects, which acts not only as guanidine hydrochloride but also as alkali. The active sites of CK have more flexibility than the whole enzyme conformation. Monomeric and dimeric molten globule-like states of CK were formed by the salt inducing in 160 and 500 mM Arg H2O solutions, respectively. The molten globule-like states indicate that monomeric and dimeric intermediates exist during CK folding. Furthermore, these results also proved the orderly folding model of CK.



Keywords and Phrases

Animal Tissue; Concentration (parameters); Controlled Study; Dissociation; Enzyme Active Site; Enzyme Activity; Enzyme Conformation; Enzyme Denaturation; Enzyme Inactivation; Enzyme Kinetics; Fluorescence; Gel Permeation Chromatography; Measurement; Muscle; Nonhuman; Polyacrylamide Gel Electrophoresis; Priority Journal; Protein Folding; Rabbit; Animal; Chemistry; Dimerization; Enzymology; Gel Chromatography; Kinetics; Methodology; Muscle; PH; Protein Denaturation; Spectrofluorometry; Ultraviolet Spectrophotometry; Oryctolagus Cuniculus; Alkali; Arginine; Creatine Kinase; Dimer; Guanidine; Monomer; Sodium Chloride; Water; Creatine Kinase MM; Isoenzyme; Potassium Chloride

International Standard Serial Number (ISSN)


Document Type

Article - Journal

Document Version


File Type





© 2003 Elsevier, All rights reserved.