Enhancement of ionic current and charge movement by coexpression of calcium channel beta 1A subunit with alpha 1C subunit in a human embryonic kidney cell line.
1. Coexpression of the beta subunit with the alpha 1C subunit of the cardiac L-type Ca2+ channel has been shown to increase ionic current. To examine the mechanism of this increase, ionic and gating currents were measured in transiently transfected HEK293 cells. 2. Beta 1A subunit coexpression increased the maximal whole-cell conductance (Gmax) measured in 10 mM Ba2+ from 91 +/- 11 to 833 +/- 107 pS pF-1 without a change in the voltage dependence of activation (V1/2: -6.1 +/- 1.1 and -6.6 +/- 0.9 mV, respectively). 3. Gating currents were smaller in cells expressing only the alpha 1C subunit (only four out of eleven cells exhibited gating currents above the limits of detection, whereas eight out of eight beta 1A coexpressing cells had measurable gating currents). The gating currents were integrated to measure the intramembrane charge movement (Q). The ON charge movement (Qon) could be described by a Boltzmann distribution reaching a maximal value of Qon,max. 4. The mean ratio of Gmax: Qon,max increased from 99 +/- 6 to 243 +/- 30 pS fC-1 with beta 1A coexpression, demonstrating that the beta 1A subunit changes the gating of alpha 1C channels to favour the opening of the channels. However, this 2.5-fold change in the Gmax: Qon,max ratio explains less than half of the 9.2-fold increase in Gmax with beta 1A subunit coexpression. The major effect is due to a 3.7-fold increase in Qon,max, demonstrating that beta 1A subunit coexpression increases the number of functional surface membrane channels.
Kamp
T J
TJ
Department of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA.
Pérez-García
M T
MT
Marban
E
E
D000200:Action Potentials; D000818:Animals; D015220:Calcium Channels / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D006801:Humans; D015640:Ion Channel Gating; D007668:Kidney / Q000502:physiology*; D051379:Mice; D018408:Patch-Clamp Techniques; D014162:Transfection
D015220:Calcium Channels