Ca(2+)-activated K+ channel and the activation of Ca2+ influx in vanadate-treated red blood cells.

The mechanism by which K+ inhibits vanadate-induced 45Ca2+ influx by human red blood cells (RBC) was studied using several independent approaches. The following results were found: 1. The inhibitory effect of K+ was absent when RBC were loaded with a Ca(2+)-chelator. This treatment at the same time inhibited the vanadate-induced K+ efflux, and the membrane hyperpolarization induced by Ca2+ in vanadate-treated cells. 2. The potency of K+, Rb+, and Cs+ to inhibit vanadate-induced Ca2+ influx corresponded to their ability to depolarize the RBC membrane via the Ca(2+)-activated K+ channel (K(Ca)). 3. Inhibition of the vanadate-induced 45Ca2+ influx by a protonophore proceeded in parallel with the inhibition of the vanadate-plus-Ca(2+)-induced membrane hyperpolarization. 4. Valinomycin in part released the inhibition of the vanadate-induced Ca2+ influx by known K(Ca) inhibitors (quinine, oligomycin, 4-aminopyridine) but not by inhibitors of the Ca2+ channel (Cu2+, HS-reagents, organic Ca2+ channel blockers). 5. K+ did not inhibit the vanadate-induced Ca2+ influx in dog RBC which have K(Ca) but no transmembrane K+ gradient. The inhibition of the vanadate-induced Ca2+ influx by external K+ appears to be due to the elimination of the electrical component of the Ca(2+)-motive force imposed by opening of the K(Ca). This implies that the Ca2+ carrier mediating the influx of Ca2+ in the presence of vanadate is of uniport type, and that the activity of K(Ca) may serve as a supporting element for Ca2+ influx.