Unidirectional K+ fluxes through recombinant Shaker potassium channels expressed in single Xenopus oocytes

We describe a method to evaluate the ratio of ionic fluxes through recombinant channels expressed in a single Xenopus oocyte. A potassium channel encoded by the Drosophila Shaker gene tested by this method exhibited flux ratios far from those expected for independent ion movement. At a fixed extracellular concentration of 25 mM K+, this channel showed single-file diffusion with an Ussing flux-ratio exponent, n', of 3.4 at a membrane potential of -30 mV. There was an apparent, small voltage dependence of this parameter with n' values of 2.4 at -15 and -5 mV. These results indicate that the pore in these channels can simultaneously accommodate at least four K+ ions. If each of these K+ ions is in contact with two water molecules, the minimum length of the pore is 24 A.