Endogenous KCNE subunits govern Kv2.1 K+ channel activation kinetics in Xenopus oocyte studies.
Kv2.1 is a voltage-gated potassium (Kv) channel that generates delayed rectifier currents in mammalian heart and brain. The biophysical properties of Kv2.1 and other ion channels have been characterized by functional expression in heterologous systems, and most commonly in Xenopus laevis oocytes. A number of previous oocyte-based studies of mammalian potassium channels have revealed expression-level-dependent changes in channel properties, leading to the suggestion that endogenous oocyte factors regulate channel gating. Here, we show that endogenous oocyte potassium channel KCNE ancillary subunits xMinK and xMiRP2 slow the activation of oocyte-expressed mammalian Kv2.1 channels two-to-fourfold. This produces a sigmoidal relationship between Kv2.1 current density and activation rate in oocyte-based two-electrode voltage clamp studies. The effect of endogenous xMiRP2 and xMinK on Kv2.1 activation is diluted at high Kv2.1 expression levels, or by RNAi knockdown of either endogenous subunit. RNAi knockdown of both xMiRP2 and xMinK eliminates the correlation between Kv2.1 expression level and activation kinetics. The data demonstrate a molecular basis for expression-level-dependent changes in Kv channel gating observed in heterologous expression studies.
Gordon
Earl
E
Greenberg Division of Cardiology, Department of Medicine, Cornell University, Weill Medical College, New York 10021, USA.
Roepke
Torsten K
TK
Abbott
Geoffrey W
GW
D000818:Animals; D003001:Cloning, Molecular; D005260:Female; D015640:Ion Channel Gating / Q000502:physiology*; D007700:Kinetics; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D024642:Potassium Channels, Voltage-Gated / Q000502:physiology*; D021122:Protein Subunits / Q000502:physiology; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D051669:Shab Potassium Channels / Q000502:physiology*; D029867:Xenopus Proteins / Q000502:physiology*; D014982:Xenopus laevis
D024642:Potassium Channels, Voltage-Gated; D021122:Protein Subunits; D034741:RNA, Small Interfering; D051669:Shab Potassium Channels; D029867:Xenopus Proteins