title: Action potential clamp and mefloquine sensitivity of recombinant 'I KS' channels incorporating the V307L KCNQ1 mutation.
abstract: The slow delayed rectifier potassium current, 'I(Ks)', contributes to repolarisation of cardiac ventricular action potentials and thereby to the duration of the QT interval of the electrocardiogram. Mutations to I(Ks) channel subunits occur in clinically significant cardiac repolarisation disorders. The short QT syndrome (SQTS) is associated with accelerated ventricular repolarisation and with an increased risk of arrhythmia and sudden death. The SQT2 variant of the SQTS has been linked to a gain-of-function amino-acid substitution (V307L) in the KCNQ1-encoded I(Ks) channel alpha-subunit. This study reports the first action potential (AP) voltage-clamp comparison between wild-type (WT) and V307L KCNQ1 (co-expressed with KCNE1 to recapitulate I(Ks)) and identifies an effective pharmacological inhibitor of recombinant 'I(Ks)' channels incorporating the V307L KCNQ1 mutation. Perforated-patch voltage-clamp recordings at 37 degrees C of whole-cell current carried by co-expressed KCNQ1 and KCNE1 showed a marked (-36 mV) shift in half-maximal activation for V307L compared to WT KCNQ1; a significant slowing of current deactivation was also observed. Under AP clamp, peak repolarising current was significantly augmented for V307L KCNQ1 compared to WT KCNQ1 for both ventricular and atrial AP commands, consistent with an ability of the V307L mutation to increase repolarising I(Ks) in both regions. The quinoline agent mefloquine inhibited WT KCNQ1+KCNE1 with an IC(50) of 3.4 muM compared to 3.3 muM for V307L KCNQ1+KCNE1 (P >0.05). This establishes mefloquine as an effective inhibitor of recombinant 'I(Ks)' channels incorporating this SQT2 KCNQ1 mutation.
authors.affiliation: Department of Physiology and Pharmacology and Cardiovascular Research Laboratories, School of Medical Sciences, University Walk, University of Bristol, UK.
authors.forename: A
authors.initials: A
authors.lastname: El Harchi
authors.forename: M J
authors.initials: MJ
authors.lastname: McPate
authors.forename: Y H
authors.initials: YH
authors.lastname: Zhang
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.forename: J C
authors.initials: JC
authors.lastname: Hancox
mesh_terms: D000200:Action Potentials / Q000187:drug effects*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004562:Electrocardiography; D004594:Electrophysiology; D020128:Inhibitory Concentration 50; D051657:KCNQ1 Potassium Channel / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015767:Mefloquine / Q000008:administration & dosage / Q000494:pharmacology*; D009154:Mutation; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000008:administration & dosage / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D051657:KCNQ1 Potassium Channel; D026902:Potassium Channel Blockers; D024642:Potassium Channels, Voltage-Gated; D015767:Mefloquine