Ca, Mg-ATPase activity of permeabilised rat heart cells and its functional coupling to oxidative phosphorylation of the cells.
Isolated rat heart cells permeabilised by digitonin were examined as an experimental model to study heart bioenergetics. The cells showed good indices of oxidative phosphorylation (acceptor control ratio about 8 with pyruvate plus malate). The adenosine triphosphatase activity detected in the cells was high and was calcium dependent (optimum [free calcium] about 400 nmol.litre-1); magnesium was necessary for its full activity. Double reciprocal plot l/v vs 1/[free calcium] at physiological free calcium concentrations was linear, thus showing free calcium to be a substrate for the adenosine triphosphatase (Km for calcium about 149 nmol.litre-1). Double reciprocal plot 1/v vs 1/[ATP] was also linear, thus showing that the adenosine triphosphatase activity could be ascribed to a single enzyme. Oxidative phosphorylation and the ATPase activity of the cells appeared to be functionally coupled. This was manifested by apparent preference by oxidative phosphorylation for adenosine diphosphate supplied by the adenosine triphosphatase activity (Km 45 mumol.litre-1) to external adenosine diphosphate (Km 152 mumol.litre-1; p less than 0.02). Apparent preference by the adenosine triphosphatase activity for adenosine triphosphate supplied by mitochondria (Km 74 mumol.litre-1) to external adenosine triphosphate (Km 169 mumol.litre-1) was also manifested by a significant difference in Km values (p less than 0.05).
Kümmel
L
L
Laboratory for Endocrinology and Metabolism, Charles University, Faculty of Medicine, Prague, Czechoslovakia.
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000187:drug effects; D004072:Digitonin / Q000493:pharmacokinetics; D006321:Heart / Q000187:drug effects; D066298:In Vitro Techniques; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology / Q000201:enzymology*; D010085:Oxidative Phosphorylation; D010101:Oxygen Consumption / Q000187:drug effects; D051381:Rats
D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D008274:Magnesium; D004072:Digitonin; D002118:Calcium