Yu, Chih-ChiehKo, Jum-SukAi, TomohikoTsai, Wen-ChinChen, ZhenhuiRubart, MichaelVatta, MatteoEverett, Thomas H.George, Alfred L.Chen, Peng-Sheng2018-02-202018-02-202016-08Yu, C.-C., Ko, J.-S., Ai, T., Tsai, W.-C., Chen, Z., Rubart, M., … Chen, P.-S. (2016). Arrhythmogenic Calmodulin Mutations Impede Activation of Small-conductance Calcium-Activated Potassium Current. Heart Rhythm : The Official Journal of the Heart Rhythm Society, 13(8), 1716–1723. https://doi.org/10.1016/j.hrthm.2016.05.0091547-5271https://hdl.handle.net/1805/15249Background Apamin sensitive small-conductance Ca2+-activated K+ (SK) channels are gated by intracellular Ca2+ through a constitutive interaction with calmodulin. Objective We hypothesize that arrhythmogenic human calmodulin mutations impede activation of SK channels. Methods We studied 5 previously published calmodulin mutations (N54I, N98S, D96V, D130G and F90L). Plasmids encoding either wild type (WT) or mutant calmodulin were transiently transfected into human embryonic kidney (HEK) 293 cells that stably express SK2 channels (SK2 Cells). Whole-cell voltage-clamp recording was used to determine apamin-sensitive current (IKAS) densities. We also performed optical mapping studies in normal murine hearts to determine the effects of apamin in hearts with (N=7) or without (N=3) pretreatment with sea anemone toxin (ATX II). Results SK2 cells transfected with WT calmodulin exhibited IKAS density (in pA/pF) of 33.6 [31.4;36.5] (median and confidence interval 25%-75%), significantly higher than that observed for cells transfected with N54I (17.0 [14.0;27.7], p=0.016), F90L (22.6 [20.3;24.3], p=0.011), D96V (13.0 [10.9;15.8], p=0.003), N98S (13.7 [8.8;20.4], p=0.005) and D130G (17.6 [13.8;24.6], p=0.003). The reduction of SK2 current was not associated with a decrease in membrane protein expression or intracellular distribution of the channel protein. Apamin increased the ventricular APD80 (from 79.6 ms [63.4-93.3] to 121.8 ms [97.9-127.2], p=0.010) in hearts pre-treated with ATX-II but not in control hearts. Conclusion Human arrhythmogenic calmodulin mutations impede the activation of SK2 channels in HEK 293 cells.en-USPublisher PolicyArrhythmogenicCalmodulin MutationsCalcium ActivatedActivation of Small-conductanceArticle