The magnesium dependence of sodium-pump-mediated sodium-potassium and sodium-sodium exchange in intact human red cells

Abstract

1. The magnesium content of human red blood cells was controlled by varying the magnesium concentration in the medium in the presence of the ionophore A23187. The new magnesium levels attained were very stable, which allowed the magnesium dependence of the sodium pump to be investigated.2. The effects of magnesium were shown to occur at the inner surface of the red cell membrane for the range of magnesium concentrations tested (10(-7) to 6 x 10(-3)m).3. At intracellular ionized magnesium concentrations below 0.8 mm the activation of ouabain-sensitive sodium-potassium exchange by internal ionized magnesium could be resolved into two or three components: (a) a small component, about 5% of the maximum flux, which is apparently independent of the ionized magnesium concentration below 2 mum, (b) a saturating component with a K((1/2)) of between 30 and 45 mum, and possibly (c) a component which increases linearly with ionized magnesium concentration and which only becomes apparent at concentrations above 0.1 mm.4. At intracellular ionized magnesium concentrations below 0.8 mm, activation of ouabain-sensitive sodium-sodium exchange by internal ionized magnesium could be resolved into two components: (a) a small component, about 6% of the maximal flux, which is apparently independent of the ionized magnesium concentration below 2 mum, and (b) a saturating component with a K((1/2)) of about 9 mum. At ionized magnesium concentrations between about 0.2 and 0.8 mm the rate of sodium-sodium exchange remained constant at the maximal level.5. The intracellular concentration of ATP decreased and the ADP concentration increased as the magnesium content of the cells was reduced from the normal level. A small increase in ATP and a small decrease in ADP was seen when the magnesium content was increased above the normal level. The variation in the ATP: ADP ratio from 2.5 at very low magnesium levels to about 6 at normal magnesium levels can account, at least in part, for the different K((1/2)) values of sodium-potassium and sodium-sodium exchange.6. When the concentration of ionized magnesium was increased above about 0.8 mm both sodium-potassium and sodium-sodium exchange were inhibited. Sodium-sodium exchange was more strongly inhibited than sodium-potassium exchange.7. The possible sites of action of magnesium in the sodium pump cycle are discussed.