The impermeant ion methylammonium blocks K+ and NH4+ currents through KAT1 channel differently: evidence for ion interaction in channel permeation

The permeation properties of KAT1, an inward rectifying potassium channel from plant cells, were investigated with different ions in the external medium. With either K+, NH4+ or methylammonium (MA) in the external solution, the channel, expressed in Xenopus oocytes, appeared permeable to K+ and, to a lesser extent, to NH4+ but not to the slightly bigger, methylated analogue of NH4+, MA. Substituting NH4+ for K+ shifted the voltage dependency of channel activation further negative and hastened activation kinetics. This suggests that channel operation depends on the transported substrate. In mixed solution (50 mM K+, 50 mM MA) MA inhibited K+ currnt in a voltage-independent manner. The maximum block did not exceed 50% of the K+ current. In contrast, when NH4+ was the permeant ion (50 mM NH4+, 50 mM MA) MA caused a voltage-dependent, slowly developing open channel block, achieving complete inhibition at very negative voltages. The latter block could be partially overcome by the addition of K+ in the external solution. The data support a model in which ions, after entering the channel pore, compete with different affinities for binding sites on their permeation pathway.