Studies on the determination of Na-K-ATPase in red blood cell membranes.
The present paper is concerned with Na-K-ATPase. In the first part it deals with the effect of such factors as Na+, K+, Mg2+, ATP, pH on activity investigated using a soluble Na-K-ATPase from porcine cerebral cortex. In the second part a method for assaying red blood cell Na-K-ATPase is described. This method consists of specific steps for washing and preparation of red cell membranes, pretreatment of Na-K-ATPase with sodium dodecyl sulfate (SDS) and bovine serum albumin (BSA) and determination of enzymatic released inorganic phosphate. Na-K-ATPase prepared and treated in this way shows a higher activity than the conventionally prepared form of enzyme, as latent enzyme sites are exposed by the SDS treatment. The kinetic behaviour of Na-K-ATPase prepared and assayed by using this method is characterized and compared with that of soluble Na-K-ATPase.
Eder
K
K
Institut für Ernährungsphysiologie, Technische Universität München-Weihenstephan, Freising, F.R.G.
Reichlmayr-Lais
A M
AM
Kirchgessner
M
M
D000818:Animals; D002540:Cerebral Cortex / Q000201:enzymology; D004910:Erythrocyte Membrane / Q000201:enzymology*; D005260:Female; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D000254:Sodium-Potassium-Exchanging ATPase / Q000097:blood*; D012995:Solubility; D013552:Swine
D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D000254:Sodium-Potassium-Exchanging ATPase