title: Interaction of creatine kinase with phosphorylating rabbit heart mitochondria and mitoplasts.
abstract: This paper demonstrates that the mitochondrial isoenzyme of creatine kinase (CKm) can be solubilized from rabbit heart mitochondria, the outer membrane of which has been removed or at least broken by a digitonin treatment or a short hypotonic exposure, but which has retained an important part of the capacity to phosphorylate ADP. Phosphate, ADP, or ATP, at concentrations which are used to study oxidative phosphorylation and creatine phosphate synthesis, solubilize CKm; the same is true with MgCl2 and KCl. The effect of adenine nucleotides does not seem to be due to their interaction with the adenine nucleotide translocase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that CKm is the main protein released in the described conditions; however, it does not amount to more than 1% of the total protein content of the mitoplasts. When the apparent Km for ATP of CKm was estimated by measuring creatine phosphate synthesis, the values obtained using water-treated mitochondria (0.21 mM) were slightly higher than those of intact mitochondria (0.12 mM) but the difference was not significant. In the former preparation 77% of CKm was in a soluble state. If we can extrapolate these results to intact mitochondria and suppose that in this case a fraction of CKm is also soluble in the intermembrane space, this does not support the theory of functional association between CKm and the adenine nucleotide translocase.
mesh_terms: D000227:Adenine Nucleotides / Q000494:pharmacology; D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism*; D004072:Digitonin; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008929:Mitochondria, Heart / Q000378:metabolism* / Q000648:ultrastructure; D010725:Phosphocreatine / Q000096:biosynthesis; D010766:Phosphorylation; D011817:Rabbits; D012995:Solubility; D014867:Water
chemical_list: D000227:Adenine Nucleotides; D010725:Phosphocreatine; D014867:Water; D003402:Creatine Kinase; D004072:Digitonin