Inhibition of human red cell sodium and potassium transport by divalent cations

The influx and efflux of Na and K across the human red cell membrane by the bumetanide-sensitive (Na-K co-transport) and residual (ouabain- and bumetanide-insensitive) routes were inhibited by increasing concentrations of external Mg. Ca, Sr, Ba, Mn and Co also inhibited bumetanide-sensitive and residual K transport. External Mg inhibited choline uptake and the Na-dependent fractions of L-alanine and L-serine uptakes. External Mg reduced the maximal rate (app. Vmax) but not the affinity (app. Km) of the bumetanide-sensitive K and Na influxes when they were measured as functions of external K and Na respectively. The inhibitory effect of Mg was not due to a small reduction in zeta potential since much larger reductions in zeta potential produced by neuraminidase did not affect transport. Internal Mg stimulated the ouabain-sensitive K influx but inhibited the co-transport and residual components of K influx. Bumetanide was a poor co-transport inhibitor in red cells pre-treated with A23187 and EDTA. It was concluded that the inhibitory effects of external Mg were probably not due to changes in the ionic composition of the diffuse double layer adjacent to the cell membrane. Mg and other divalent cations should not be used as 'inert' ionic substitutes in human red cell Na and K transport studies.