title: Partial purification of the ouabain-binding component and of Na,K-ATPase from human red cell membranes.
abstract: Ghosts of human red cells were incubated with tritiated ouabain in the presence of ATP, Mg, and Na, conditions under which ouabain binds with high specificity to Na:K transport sites. The labeled membranes were solubilized with sodium dodecyl sulfate and dialyzed. Sodium dodecyl sulfate solubilizes most of the membrane protein and leaves most of the tritiated-ouabain bound to a solubilized component. Solubilized Na,K-ATPase could also be obtained after dialysis. The solubilized membranes were centrifuged in a sucrose density gradient. The ouabain-membrane complex and the Na,K-ATPase sedimented faster than the bulk of the protein. The ouabain-membrane complex and the Na,K-ATPase appeared to be identical and were purified about eightfold relative to the starting material. These results represent a step toward the isolation and characterization of the cation transport mechanism in red cell membranes.
mesh_terms: D000251:Adenosine Triphosphatases / Q000302:isolation & purification*; D001693:Biological Transport, Active; D002462:Cell Membrane / Q000032:analysis* / Q000201:enzymology* / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D004912:Erythrocytes / Q000166:cytology* / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D010042:Ouabain; D011188:Potassium; D012964:Sodium; D012995:Solubility; D013501:Surface-Active Agents; D014316:Tritium
chemical_list: D013501:Surface-Active Agents; D014316:Tritium; D010042:Ouabain; D012964:Sodium; D000251:Adenosine Triphosphatases; D011188:Potassium