Antigenic differences in (Na+, K+)-ATPase preparations isolated from various organs and species.
Antisera to purified (Na+, K+)-ATPase raised in rabbits and in sheep were purified by an absorption procedure employing purified canine kidney (Na+, K+)-ATPase. The antibodies were fractionated into two components, one which inhibited catalytic activity, and a second which inhibited ouabain binding. Under certain conditions, the fraction that inhibited ouabain binding also inhibited catalytic activity, and the effectiveness of both was dependent to some extent on the ligands present in the incubation medium. Thus, both antibody fractions appeared to detect conformations of the enzyme that depended upon ligand-induced perturbations. When the antibody raised against catalytic activity was incubated with erythrocyte membrane fragments, an inhibition of the (Na+, K+)-ATPase occurred, but only minimal or no effect on potassium influx or on digoxin-induced inhibition of potassium flux in intact erythrocytes was noted. In a similar experiment, however, the antibody against ouabain binding significantly inhibited potassium influx, suggesting specificity in terms of the macromolecular surfaces of the pump which were exposed to the external medium. We concluded that there may be organ and species differences among (Na+, K+)-ATPase preparations. Antibodies prepared in rabbits and sheep were fractionated by absorption to dog brain enzyme. Both the antibody fraction which bound to the brain enzyme and that which did not bind inhibited the dog kidney (Na+, K+)-ATPase, but only the former inhibited dog brain (Na+, K+)-ATPase. When the two fractions were recombined, inhibition was restored to the extent of the unfractionated antibody.
D000251:Adenosine Triphosphatases / Q000097:blood / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D000937:Antigen-Antibody Reactions; D001693:Biological Transport, Active; D001921:Brain / Q000201:enzymology; D003429:Cross Reactions; D004077:Digoxin / Q000378:metabolism; D004285:Dogs; D004789:Enzyme Activation; D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D007122:Immunoelectrophoresis; D007668:Kidney / Q000201:enzymology; D009928:Organ Specificity; D010042:Ouabain / Q000378:metabolism; D011188:Potassium / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding; D012964:Sodium / Q000494:pharmacology; D013045:Species Specificity
D010042:Ouabain; D004077:Digoxin; D012964:Sodium; D000251:Adenosine Triphosphatases; D011188:Potassium