Abstract
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 (≥6 M) eliminated the subunit protection. Subunit interactions are also important in preserving secondary structure and forming contracted conformation at low urea concentration.
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Guo, Z., Wang, Z. & Wang, X.C. Subunit Interaction Slows the Unfolding of the N-Terminal Domain of Creatine Kinase in Urea. Biochemistry (Moscow) 67, 1388–1394 (2002). https://doi.org/10.1023/A:1021866227969
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DOI: https://doi.org/10.1023/A:1021866227969