Membrane mediated link between ion transport and metabolism in human red cells.
When 10(-6) M oubain is added to human red cell that have been incubated without glucose for two hours, there is a significant shift in the 31P nuclear magnetic resonances of both phosphate groups of cellular 2,3-diphosphoglycerate, which is not found in control cells incubated with glucose. This means that an effect induced by ouabain on the outside of the red cell membrane is transmitted through the membrane to alter the environment of an intracellular metabolite. Experiments with glycolytic cycle inhibitors have indicated that the intracellular ligand responsible for the resonance shifts is monophosphoglycerate mutase which requires 2,3-diphosphoglycerate as a cofactor for the reaction it catalyzes. To account for this finding a hypothesis is presented that the (Na+ + K+)-ATPase in human red cells is linked to monophosphoglycerate mutase through the agency of phosphoglycerate kinase. Evidence is presented for the existence of phosphoglycerate kinase/monophosphoglycerate mutase in solution. It is shown that this complex can interact with the cytoplasmic face of (Na+ + K+)-ATPase at the outside surface of inside out red cell vesicles, and that this interaction is inhibited when 10(-6) M ouabain is contained within the vesicle. Neither monophosphoglycerate mutase nor phosphoglycerate kinase is significantly bound to the inside surface of the intact human red cell, but glyceraldehyde 3-phosphate dehydrogenase is; it is shown that this enzyme also interacts with the cytoplasmic face of the (Na+ + K+)-ATPase and that the interaction is inhibited by 10(-6) M ouabain.
D001693:Biological Transport, Active; D001786:Blood Glucose / Q000378:metabolism; D004163:Diphosphoglyceric Acids / Q000097:blood; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D005459:Fluorides / Q000494:pharmacology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000097:blood; D006019:Glycolysis; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000097:blood*; D012413:Rubidium / Q000097:blood*; D013431:Sulfates / Q000494:pharmacology
D001786:Blood Glucose; D004163:Diphosphoglyceric Acids; D013431:Sulfates; D010042:Ouabain; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D012413:Rubidium; D005459:Fluorides; D011188:Potassium