Purification of mitochondrial creatine kinase. Biochemical and immunological characterization.
Mitochondrial creatine kinase was purified from canine myocardium. The preparation exhibited a positively charged isoenzyme free of other creatine kinase isoenzymes and on sodium dodecyl sulfate gel exhibited a single protein band. Amino acid composition showed mitochondrial creatine kinase to be different from that of MM or BB creatine kinase and did not hybridize with the M or B subunits of the cytosolic forms. Antiserum was developed to mitochondrial creatine kinase which did not cross-react with cytosolic creatine kinases. Antiserum to cytosolic creatine kinase exhibited no reaction to mitochondrial creatine kinase. Utilizing the specific antiserum, a radioimmunoassay was developed for the specific detection of mitochondrial creatine kinase. Thus, mitochondrial creatine kinase was purified and shown to be comprised of a unique subunit which is biochemically and immunologically distinct from the cytosolic creatine kinases.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001921:Brain / Q000201:enzymology; D003402:Creatine Kinase / Q000276:immunology / Q000302:isolation & purification*; D003600:Cytosol / Q000201:enzymology; D004285:Dogs; D007106:Immune Sera; D007118:Immunoassay; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007527:Isoenzymes; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology*; D008929:Mitochondria, Heart / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology
D000596:Amino Acids; D007106:Immune Sera; D007527:Isoenzymes; D046911:Macromolecular Substances; D003402:Creatine Kinase