Studies on the mechanism of inhibition of the red cell metabolism by cardiac glycosides.
The purpose of this work was to test the previously suggested hypothesis that the inhibitory effect of ouabain on lactate production in human red cells is due to an interaction between phosphoglycerate kinase and (Na+ + k+)-activated adenosine triphosphatase (Na+,K+ATPase). An antibody to red cell phosphoglycerate kingase caused complete inhibition of the purified enzyme, whereas a portion of the phophoglycerate kinase activity of the red cell membranes was resistant to the antibody. When increasing amounts of the purified enzyme were added to the membranes, the antibody-resistant portion of the activity increased. The effects of the antibody and ouabain on lactate production from fructose-6,6-diphosphate in red cell hemolysates were studied. Ouabain, at a maximally effective concentration, produced about 30% inhibition of lactate formation. This value was doubled in the presence of the antibody. Red cell membranes, and rat brain Na+,K+-ATPase, did not catalyze the hydrolysis of 1,3-diphosphoglycerate. Ouabain did not affect the reactions of the Rapport-Luebering pathway of the red cells. These findings provide further support for the view that in red cells a membrane pool of phosphoglycerate kinase is oriented in the vicinity of Na+,K+-ATPase in a way that the product of each enzyme may be used as the immediate substrate of the other and that ouabain inhibits glycolysis by removing the regulatory effect of Na+,K+-ATPase on that portion of glycolysis which is channeled through this pool of phosphoglycerate kinase.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001666:Binding Sites, Antibody; D002463:Cell Membrane Permeability / Q000187:drug effects; D004163:Diphosphoglyceric Acids / Q000378:metabolism; D004347:Drug Interactions; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D006019:Glycolysis / Q000187:drug effects; D006801:Humans; D007773:Lactates / Q000096:biosynthesis; D009943:Organophosphorus Compounds / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology*; D010735:Phosphoglycerate Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D012964:Sodium / Q000378:metabolism
D004163:Diphosphoglyceric Acids; D007773:Lactates; D009943:Organophosphorus Compounds; D010042:Ouabain; D012964:Sodium; D010735:Phosphoglycerate Kinase; D000251:Adenosine Triphosphatases; D011188:Potassium