Differential sensitivity of chicken MM-creatine kinase to trypsin and proteinase-K. Int J Biochem 1985;17(3):309-18
Date
01/01/1985Pubmed ID
3891449DOI
10.1016/0020-711x(85)90205-8Scopus ID
2-s2.0-0021840560 (requires institutional sign-in at Scopus site) 11 CitationsAbstract
Under several conditions of SDS-PAGE, the chicken MM-creatine kinase (MM-CK) monomer migrated as a approximately 50,000 dalton polypeptide, approx 25% larger than usually reported. Characterization by sedimentation equilibrium indicated that the anomalous molecular weight was an artifact of electrophoresis. Digestion with trypsin caused only moderate reductions in CK activity, despite extensive degradation of the denatured enzyme revealed by SDS-PAGE. Characterization of trypsinized MM-CK under non-denaturing conditions of electrophoresis and HPLC revealed no fragmentation of the native enzyme, suggesting that MM-CK quaternary structure was maintained despite extensive tryptic nicking. In contrast, much lower concentrations of proteinase-K generated only a single fragment in SDS-PAGE while causing a nearly total loss of enzyme activity.
Author List
Lough J, Wrenn DS, Miziorko HM, Auer HEAuthor
John W. Lough PhD Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino AcidsAnimals
Chickens
Chromatography, High Pressure Liquid
Creatine Kinase
Electrophoresis, Polyacrylamide Gel
Endopeptidase K
Endopeptidases
Isoenzymes
Trypsin