Combination of the electrogenic ionophores, valinomycin and CCCP, can lead to non-electrogenic K+/H+ exchange on bilayer lipid membranes.

The method of pH shift measuring by means of a pH microelectrode was applied to measure hydrogen ion fluxes across a planar bilayer lipid membrane (BLM) in the presence of the potassium ion ionophore, valinomycin, and a protonophore, carbonylcyanide m-chlorophenylhydrazone (CCCP), under conditions of the voltage clamp. The voltage dependence of the flux was determined to be in the range of +/- 150 mV under the conditions of both symmetrical KCl as well as a KCl gradient across the BLM. Surprisingly, at a clamped zero voltage on BLM a significant hydrogen ion flux was observed in the presence of a KCL gradient and both valinomycin and CCCP. This finding was interpreted as a result of induction of non-electrogenic K+/H+ exchange in the presence of valinomycin and CCCP, presumably through the formation of electrically neutral complexes of these two ionophores and K+ (H+) ions: valinomycin-K(+)-CCCP- and/or possibly valinomycin-CCCP(-)-H+.