Subunit interaction slows the unfolding of the N-terminal domain of creatine kinase in urea.
Fluorescence emission intensity changes with two different excitation wavelengths were used to measure the unfolding rate constants of different domains of muscle type creatine kinase (CK-MM) according to the heterogeneity of aromatic amino acid distributions in the crystal structure of CK-MM. The results were compared with those of brain type creatine kinase (CK-BB) and dithio-bis(succinimidyl propionate) cross-linked CK-MM. CK-BB differed greatly in its distribution of aromatic amino acids in each domain and the unfolding process of cross-linked CK-MM was not accompanied by the dissociation of the dimer. The N-terminal domain of CK-MM was shown to be well protected by subunit interaction during the unfolding of CK-MM in 4 M urea. Dissociating the CK dimer in high urea concentration (> or = 6 M) eliminated the subunit protection. Subunit interactions are also important in preserving secondary structure and forming contracted conformation at low urea concentration.