Mechanisms for Kir channel inhibition by quinacrine: Acute pore block of Kir2.x channels and interference in PIP 2 interaction with Kir2.x and Kir6.2 channels
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Cardiac inward rectifier potassium currents determine the resting membrane potential and contribute repolarization capacity during phase 3 repolarization. Quinacrine is a cationic amphiphilic drug. In this work, the effects of quinacrine were studied on cardiac Kir channels expressed in HEK 293 cells and on the inward rectifier potassium currents, I K1 and I KATP, in cardiac myocytes. We found that quinacrine differentially inhibited Kir channels, Kir6.2∈∼∈Kir2.3∈>∈Kir2.1. In addition, we found in cardiac myocytes that quinacrine inhibited I KATP∈>∈I K1. We presented evidence that quinacrine displays a double action towards strong inward rectifier Kir2.x channels, i.e., direct pore block and interference in phosphatidylinositol 4,5-bisphosphate, PIP 2-Kir channel interaction. Pore block is evident in Kir2.1 and 2.3 channels as rapid block; channel block involves residues E224 and E299 facing the cytoplasmic pore of Kir2.1. The interference of the drug with the interaction of Kir2.x and Kir6.2/SUR2A channels and PIP 2 is suggested from four sources of evidence: (1) Slow onset of current block when quinacrine is applied from either the inside or the outside of the channel. (2) Mutation of Kir2.3(I213L) and mutation of Kir6.2(C166S) increase their affinity for PIP 2 and lowers its sensitivity for quinacrine. (3) Mutations of Kir2.1(L222I and K182Q) which decreased its affinity for PIP 2 increased its sensitivity for quinacrine. (4) Co-application of quinacrine with PIP 2 lowers quinacrine-mediated current inhibition. In conclusion, our data demonstrate how an old drug provides insight into a dual a blocking mechanism of Kir carried inward rectifier channels. © 2011 Springer-Verlag.
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Inward rectifier potassium channels; Kir2.1; Kir2.3; Kir6.2; Phosphatidylinositol 4,5-bisphosphate, PIP 2; Quinacrine inwardly rectifying potassium channel; inwardly rectifying potassium channel subunit Kir2.x; inwardly rectifying potassium channel subunit Kir6.2; mepacrine; phosphatidylinositol 4,5 bisphosphate; unclassified drug; article; binding affinity; cellular distribution; channel gating; controlled study; drug mechanism; drug sensitivity; drug targeting; heart muscle cell; human; human cell; IC 50; mutational analysis; priority journal; protein expression; protein localization; protein protein interaction; residue analysis; sensitivity analysis; HEK293 Cells; Humans; Myocytes, Cardiac; Phosphatidylinositol 4,5-Diphosphate; Potassium Channels, Inwardly Rectifying; Quinacrine
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