Human erythrocyte membranes exhibit a cooperative calmodulin-dependent Ca2+-ATPase of high calcium sensitivity.
It is thought that the ionized Ca2+ concentration in the cytosol of healthy erythrocytes is in the range of 0.01--0.1 microM (ref. 1) and that this low concentration is maintained by an ATP-driven calmodulin-dependent Ca2+ pump in the plasma membrane. The Ca2+-stimulated ATPase which is the enzymatic expression of this pump varies in its calcium sensitivity between different preparations of erythrocyte ghosts, with activation generally occurring in a concentration range between approximately 1 and 10 microM Ca2+ (refs 2--5). This is a higher range of Ca2+ concentrations than might be anticipated for activation of a pump that sustains intracellular concentrations of Ca2+ below 0.1 microM, and recent reports have suggested activation in some membrane preparations at Ca2+ concentrations in the range 0.1--1.0 microM (refs 6, 7). We report here a simple method for preparing human erythrocyte membranes in 2.5 mM HEPES/1 mM EGTA at pH 7.0 (see Fig. 1 legend) in which the activation of the Ca2+-ATPase by Ca2+ and intracellular concentrations of calmodulin is highly cooperative and is complete by approximately 1 microM Ca2+. Unlike other available erythrocyte membrane preparations, the pattern of activation by Ca2+ and calmodulin is not complicated by partial resealing of the ghosts during and after isolation. We suggest that this cooperative activation of the Ca2+ pump may explain how healthy erythrocytes maintain their normal cytosol Ca2+ concentration at a threshold value at or below approximately 0.1 microM. We also note that several other calmodulin-dependent enzymes display similar cooperative activation kinetics.
D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002147:Calmodulin / Q000494:pharmacology*; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000201:enzymology*; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D002147:Calmodulin; D008565:Membrane Proteins; D000252:Calcium-Transporting ATPases; D002118:Calcium