Potassium: potassium exchange catalysed by the sodium pump in human red cells

Abstract

1. When red cells were so depleted of Na that Na:K exchange had almost ceased, the ouabain-sensitive K efflux seen in K-containing media was accompanied by an almost equal ouabain-sensitive K influx.2. This suggests that the Na pump in these cells was carrying out a one-for-one K:K exchange across the erythrocyte membrane.3. 30-40% of the (42)K efflux from resealed ghosts was sensitive to ouabain when the ghosts contained 1 mM-ATP, 2 mM orthophosphate, 10 mM-K and less than 1 mM-Na, and the suspending medium contained 10 mM-K and 0-Na, choline being the predominant cation.4. In resealed ghosts, the rate of K:K exchange saturated as internal K was increased, and was half-maximal at about 10 mM-K.5. When internal ATP was maintained with a phosphocreatine:creatine phosphokinase regenerating system, K:K exchange saturated as internal ATP was increased, and was half-maximal at about 100 muM-ATP.6. The rate of K:K exchange did not depend on whether the ADP concentration was roughly the same as the ATP concentration or very much less, suggesting that ADP did not affect the rate of K:K exchange.7. GTP, ITP and UTP were unable to substitute for ATP in supporting K:K exchange. CTP was a poor substitute.8. There was no evidence to support the hypothesis that K:K exchange is accompanied by a ouabain-sensitive hydrolysis of ATP.9. Internal Na was a strong inhibitor of ouabain-sensitive K efflux from ghosts containing 9 mM-K. 4 mM-Na was sufficient to produce 90% inhibition.10. The rate of K:K exchange depended on the orthophosphate concentration inside the ghosts (confirming Glynn, Lew & Lüthi, 1970). The curve obtained suggested that the rate was half-maximal at about 1.7 mM orthophosphate.11. These experiments suggested that inhibition by internal K is an important factor affecting the Na efflux from intact red cells. Experiments measuring Na:K exchange as a function of internal Na in low-K ghosts supported this hypothesis.12. The significance of these findings is discussed.