Gating of a pH-sensitive K_2P potassium channel by an electrostatic effect of basic sensor residues on the selectivity filter

K⁺ channels share common selectivity characteristics but exhibit a wide diversity in how they are gated open. Leak K_2P K⁺ channels TASK-2, TALK-1 and TALK-2 are gated open by extracellular alkalinization. The mechanism for this alkalinizationdependent gating has been proposed to be the neutralization of the side chain of a single arginine (lysine in TALK-2) residue near the pore of TASK-2, which occurs with the unusual pK_a of 8.0. We now corroborate this hypothesis by transplanting the TASK-2 extracellular pH (pH_o) sensor in the background of a pH_o-insensitive TASK-3 channel, which leads to the restitution of pH_o-gating. Using a concatenated channel approach, we also demonstrate that for TASK-2 to open, pH_o sensors must be neutralized in each of the two subunits forming these dimeric channels with no apparent cross-talk between the sensors. These results are consistent with adaptive biasing force analysis of K⁺ permeation using a model selectivity filter in wild-type and mutated channels. The underlying free-energy profiles confirm that either a doubly or a singly charged pH_o sensor is sufficient to abolish ion flow. Atomic detail of the associated mechanism reveals that, rather than a collapse of the pore, as proposed for other K_2P channels gated at the selectivity filter, an increased height of the energetic barriers for ion translocation accounts for channel blockade at acid pH_o. Our data, therefore, strongly suggest that a cycle of protonation/deprotonation of pH_o-sensing arginine 224 side chain gates the TASK-2 channel by electrostatically tuning the conformational stability of its selectivity filter.