Influence of L-type Ca channel alpha 2/delta-subunit on ionic and gating current in transiently transfected HEK 293 cells.
We have measured ionic and gating currents in human embryonic kidney (HEK 293) cells transiently transfected with cDNAs encoding subunits of the cardiac voltage-gated L-type Ca2+ channel. Robust recombinant ionic current and associated nonlinear charge movement could be measured over a broad voltage range without contamination by endogenous channel activity. Coexpression of the alpha 2/delta-subunit along with alpha 1- and beta 2-subunits speeded activation and deactivation kinetics and significantly increased the maximal conductance of ionic current. Charge movement was measured at voltages negative to the threshold for activation of ionic current, and gating charge could be immobilized at positive holding potentials that did not inactivate ionic current. The ratio of maximal ionic conductance to maximal charge moved remained the same in the absence or presence of the alpha 2/delta-subunit. However, the maximal amount of charge moved was increased about twofold in the presence of the alpha 2/delta-subunit. These results suggest that coexpression of the alpha 2/delta-subunit enhances the expression of functional L-type channels and, in addition, provide evidence that most of the L-type channel-associated nonlinear charge movement is caused by transitions between nonconducting states of the channel protein that precede the open and inactivated states.
Bangalore
R
R
Department of Physiology, University of Rochester School of Medicine New York 14642, USA.
Mehrke
G
G
Gingrich
K
K
Hofmann
F
F
Kass
R S
RS
D015220:Calcium Channels / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D004056:Differential Threshold; D004553:Electric Conductivity; D006801:Humans; D015640:Ion Channel Gating; D007477:Ions; D007668:Kidney / Q000166:cytology / Q000196:embryology / Q000502:physiology*; D013997:Time Factors; D014162:Transfection
D015220:Calcium Channels; D007477:Ions