Partial purification and characterization of (Na+ + K+)-ATPase from garfish olfactory nerve axon plasma membrane.
The (Na+ + K+)-ATPase of garfish olfactory nerve axon plasma membrane was purified about sixfold by treatment of the membrane with sodium dodecyl sulfate followed by sucrose density gradient centrifugation. The estimated molecular weights of the two major polypeptide components of the enzyme preparation on sodium dodecyl sulfate gels were 110,000 and 42,000 daltons, which were different from those of the corresponding peptides of rabbit kidney (Na+ + K+)-ATPase. No carbohydrate was detected in the 42,000-dalton component either by the periodic acid-Schiff reagent or by the more sensitive concanavalin A-peroxidase staining procedure. The molecular properties of the garfish (Na+ + K+)-ATPase, such as the Km for ATP, pH optimum, energies of activation, Na and K ion dependence and vanadium inhibition, were, however, similar to those of the kidney enzyme. The partially purified garfish (Na+ + K+)-ATPase was reconstituted into phospholipid vesicles by a freeze-thaw-sonication procedure. The reconstituted enzyme was found to catalyze a time and ATP dependent 22Na+ transport. The ratio of 22Na+ pumped to ATP hydrolyzed was about 1; under the same reconstitution and assay conditions, eel electroplax (Na+ + K+)-ATPase, however, gave a 22Na+ pumped to ATP hydrolyzed ratio of nearly 3.
D000818:Animals; D001369:Axons / Q000201:enzymology*; D001693:Biological Transport, Active / Q000187:drug effects; D002462:Cell Membrane; D005399:Fishes; D007700:Kinetics; D008563:Membrane Lipids / Q000302:isolation & purification; D008565:Membrane Proteins / Q000302:isolation & purification; D009832:Olfactory Nerve / Q000201:enzymology*; D010042:Ouabain / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000302:isolation & purification* / Q000378:metabolism
D008563:Membrane Lipids; D008565:Membrane Proteins; D010042:Ouabain; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase