Zinc and mercuric ions distinguish TRESK from the other two-pore-domain K+ channels

TWIK-related spinal cord K+ channel (TRESK) is the most recently cloned two-pore-domain potassium (2PK(+)) channel, regulated by the calcium/calmodulin-dependent protein phosphatase calcineurin. Functional identification of endogenous TRESK and its distinction from the other 2PK(+) channels, producing similar background K+ current, are impeded by the lack of specific inhibitors. Therefore, we searched for antagonists selective against TRESK among the mouse 2PK(+) channels by screening more than 200 substances. Mibefradil, zinc, and mercuric ions inhibited TRESK expressed in Xenopus laevis oocytes with IC50 values lower than 10 mu M. The specificity of the identified agents was determined by measuring their effects on mouse TALK-1, TASK-1, TASK-2, TASK-3, THIK-1, TRAAK, TREK-1, and TREK-2. Mibefradil failed to discriminate well among the functional 2PK(+) channels; however, Zn2+ and Hg(2+)exerted a significantly stronger inhibitory effect on TRESK than on the other channels. Sensitivity to zinc but insensitivity to ruthenium red were distinctive features of TRESK. Whereas both Zn2+ and Hg2+ were selective blockers of TRESK among the mouse 2PK(+) channels, human TRESK was resistant to Zn2+; it was blocked only by Hg2+. His132 of mouse TRESK was partly responsible for this difference. Mouse TRESK expressed in COS-7 cells was also inhibited by Zn2+ and Hg2+, and TRESK single-channel current was diminished in outsideout patches, indicating that the action of the ions was membrane-delimited, most probably targeting the channel itself. Thus, both Zn2+ and Hg2+ are expected to inhibit endogenous TRESK in isolated mouse cells, and these ions can be applied to identify the calcineurin-activated 2PK(+) channel in its natural environment.