Retarded diffusion of ADP in cardiomyocytes: possible role of mitochondrial outer membrane and creatine kinase in cellular regulation of oxidative phosphorylation.
Possible reasons for retarded intracellular diffusion of ADP were investigated. The isolated skinned cardiac fibers were used to study apparent kinetic parameters for externally added ADP in control of mitochondrial respiration. Participation of myosin-ATPase in binding of ADP within cells as it was supposed earlier (Saks, V.A., Belikova, Yu.O. and Kuznetsov, A.V. (1991) Biochim. Biophys. Acta 1074, 302-311) was completely excluded, since myosin-deprived skinned cardiac fibers ('ghosts') displayed the same kinetic parameters as intact ones (Kmapp for ADP about 300 microM). Significantly lower apparent Km values were obtained for fibers with osmotically disrupted outer mitochondrial membrane (25-35 microM), which was close to that observed for isolated heart mitochondria. The data obtained are in favor of limitation of ADP movement via anion-selective low-conductance porine channels in the outer membrane of mitochondria. It is proposed that the permeability of this membrane is controlled by some unknown intracellular factor(s). In the presence of saturating concentrations of creatine (25 mM) the apparent Km for ADP significantly decreases due to coupling of creatine kinase and oxidative phosphorylation reactions in mitochondria. This coupling is not observed in KCl medium in which mitochondrial creatine kinase is detached from the membrane. It is concluded that in the cells in-vivo ADP movement between cytoplasm and intramitochondrial space is controlled by low-conductivity anion channels in the outer membrane. Thus, the mitochondrial creatine kinase reaction coupled to the adenine nucleotide translocase is an important mechanism in control of oxidative phosphorylation in vivo due to its ability to manifold amplify these very weak ADP signals from cytoplasm.
Laboratory of Bioenergetics, Cardiology Research Center, Moscow, Russia.
V A
VA
Saks
E
E
Vasil'eva
Belikova YuO
A V
AV
Kuznetsov
S
S
Lyapina
L
L
Petrova
N A
NA
Perov
D000244:Adenosine Diphosphate / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D003401:Creatine / Q000378:metabolism; D003402:Creatine Kinase / Q000378:metabolism*; D004058:Diffusion; D007425:Intracellular Membranes / Q000378:metabolism; D008297:Male; D008929:Mitochondria, Heart / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism* / Q000648:ultrastructure; D010085:Oxidative Phosphorylation; D018272:Porins; D011189:Potassium Chloride; D051381:Rats; D017208:Rats, Wistar
D001425:Bacterial Outer Membrane Proteins; D018272:Porins; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D003402:Creatine Kinase; D003401:Creatine