Asymmetry of Na-K-Cl cotransport in human erythrocytes

The Na-K-Cl cotransport system in human erythrocytes was studied by measuring net influxes and effluxes of Na and K. The influx of K was shown to be stimulated by Na and the influx of Na was stimulated by K, satisfying the fundamental criterion of cotransport. In addition, these mutually stimulating cation influxes had a stoichiometry of 1:1 and were entirely inhibited by furosemide; these results are also consistent with cotransport. Furthermore, the mutually stimulating influxes were entirely dependent on Cl, since they were abolished when nitrate was substituted for Cl. In contrast, cotransport, defined by mutual dependence of fluxes, was not detected in the outward direction over a range of cellular Na and K concentrations from 0 to 50 mmol/l cells. The cotransport pathway did, however, appear to mediate a Na-stimulated K efflux (but no K-stimulated Na efflux), and furosemide-inhibitable effluxes of both Na and K. Nitrate (but not sulfate) appeared to substitute for chloride in promoting Na-stimulated K efflux. Thus the Na-K-Cl cotransport system in human red cells is intrinsically asymmetric, and mediates coupled cation fluxes readily only in the inward direction.