Modeling of the adrenergic response of the human I^sub Ks^ current (hKCNQ1/hKCNE1) stably expressed in HEK-293 cells

Stable coexpression of human (h)KCNQ1 and hKCNE1 in human embryonic kidney (HEK)-293 cells reconstitutes a nativelike slowly activating delayed rectifier ... current (HEK-I...), allowing β-adrenergic modulation of the current by stimulation of endogenous receptors in the host cell line. HEK-I... was enhanced two- to fourfold by isoproterenol (EC... = 13 nM), forskolin (10 ...M), or 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (50 ...M), indicating an intact cAMP-dependent ion channel-regulating pathway analogous to the PKA-dependent regulation observed in native cardiac myocytes. Activation kinetics of HEK-IKs were accurately fit with a novel modified second-order Hodgkin-Huxley (H-H) gating model incorporating a fast and a slow gate, each independent of each other in scale and adrenergic response, or a "heterodimer" model. Macroscopically, β-adrenergic enhancement shifted the current activation threshold to more negative potentials and accelerated activation kinetics while leaving deactivation kinetics relatively unaffected. Modeling of the current response using the H-H model indicated that observed changes in gating could be explained by modulation of the opening rate of the fast gate. Under control conditions at nearly physiological temperatures (35...C), rate-dependent accumulation of HEK-I... was observed only at pulse frequencies exceeding 3 Hz. Rate-dependent accumulation of I... at high pulsing rate had two phases, an initial staircaselike effect followed by a slower, incremental accumulation phase. These phases are readily interpreted in the context of a heterodimeric H-H model with two independent gates with differing closing rates. In the presence of isoproterenol after normalizing for its tonic effects, rate-dependent accumulation of HEK-I... appeared at lower pulse frequencies and was slightly enhanced (...25%) over control. (ProQuest: ... denotes formulae/symbols omitted.)