Once again about the functional coupling between mitochondrial creatine kinase and adenine nucleotide translocase.

Abstract

The synthesis of creatine phosphate (CP) by mitochondrial creatine kinase during oxidative phosphorylation was terminated when the mass action ratio of the creatine kinase reaction Gamma = [ADP]*[CP]/[ATP]*[Cr] became equal to the apparent equilibrium constant (K(eq)(app))of this reaction. Subsequent excess of Gamma over the K(eq)(app) was due to an increase in the ADP concentration in the medium. A comparable increase in the ADP concentration also occurred in the absence of creatine (Cr) in the incubation medium. Increase in the ADP concentration was shown to be associated with a decrease in the rate of oxidative phosphorylation and with a relative increase in the ATPase activity of mitochondria during the incubation. A low concentration of ADP (<30 micro M) and relatively high concentrations (1-6 mM) of other components of the creatine kinase reaction prevented the detection of the reverse reaction within 10 min after Gamma exceeded the K(eq)(app), but the reverse reaction became evident on more prolonged incubation. The reverse reaction was accompanied by a further increase in Gamma. Low ADP concentration in the medium was also responsible for the lack of an immediate conversion of the excess creatine phosphate added although Gamma > K(eq)(app). The findings are concluded to be in contradiction with the concept of microcompartment formation between mitochondrial creatine kinase and adenine nucleotide translocase.