The Ca2+-sensitive K+-conductance of the human red cell membrane is strongly dependent on cellular pH.

The conductance of the Ca2+-sensitive K+-channels in human red cell membranes has been determined as a function of the intracellular pH. A sudden increase in the intracellular concentration of ionized calcium was established by addition of ionophore A23187 to a suspension of cells in buffer-free, Ca2+-containing salt solution. At the various cellular pH-values cellular concentrations of ionized Ca, saturating with respect to activation of the Ca2+-sensitive K+-conductance, were obtained by the use of varied concentrations of extracellular Ca2+ and added ionophore A23187. Changes in membrane potential was monitored as CCCP-mediated changes in extracellular pH. Initial net effluxes of K+, cellular K+ contents and the K+ Nernst equilibrium potentials were calculated from flame photometric measurements. Cellular Ca-contents were determined by aid of 45Ca. With cellular Ca2+ at the saturating level with respect to activation of the K+-channel the K+-conductance calculated from these data was independent of extracellular pH and a steep function of cellular pH with a half maximal conductance of 31 microSeconds/cm2 at a cellular pH of 6.1. The K+-conductance is not a simple function of cellular pH (pHc). From pHc = 6.5 and down to pHc = 6.0 a Hill-coefficient of 2.5 was found, indicating cooperativity between at least two sites regulating the conductance. Below pHc = 6.0 an extremely high Hill-coefficient of 11 was found, probably indicating that the additional titration of the channel protein leads to an increased cooperativity. The importance, as a physiological regulatory mechanism, of a K+-conductance increasing from zero to maximal conductance within less than one unit of pH, is discussed.