Method for the preparation of human erythrocyte membrane with low basal calcium ATPase, responsive to stimulation.
A simple procedure for preparing erythrocyte membranes with low basal Ca2+ ATPase activity is described, which is stimulated several-fold by the addition of hemolysate in the incubation mixture. The cells are hemolyzed in hypotonic imidazole buffer and resulting membranes are washed with hypotonic phosphate buffer (pH 8.0) and the hemolyzing medium. The membrane preparations also have Mg2+-stimulated and Na+-K+-stimulated ATPase activities. The method allows the comparison of basal Ca2+ ATPase as well as hemolysate- or calmodulin-stimulated Ca2+ ATPase activities and thus may be useful in studying Ca2+ ATPase activity in various physiopathological conditions.
Thaker
J H
JH
D000251:Adenosine Triphosphatases / Q000097:blood; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases / Q000097:blood*; D002458:Cell Fractionation / Q000379:methods; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000201:enzymology*; D006461:Hemolysis; D006801:Humans; D000254:Sodium-Potassium-Exchanging ATPase / Q000097:blood
D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D000254:Sodium-Potassium-Exchanging ATPase