Regulation of human neuronal calcium channels by G protein betagamma subunits expressed in human embryonic kidney 293 cells.
We examined the ability of different G protein subunits to inhibit the activity of human alpha1B and alpha1E Ca2+ channels stably expressed in human embryonic kidney (HEK) 293 cells together with beta1B and alpha2Bdelta Ca2+ channel subunits. Under normal conditions, Ca2+ currents in alpha1B-expressing cells showed little facilitation after a depolarizing prepulse. However, when we overexpressed the beta2gamma2 subunits of heterotrimeric G proteins, the time course of activation of the Ca2+ currents was considerably slowed and a depolarizing prepulse produced a large facilitation of the current as well as an acceleration in its time course of activation. Similar effects were not observed when cells were transfected with constitutively active mutants of the G protein alpha subunits alpha s, alpha i1, and alpha o or with the G protein beta2 and gamma2 subunits alone. Studies carried out in cells expressing alpha1E currents showed that overexpression of beta2gamma2 subunits produced pre-pulse facilitation, although this was of lesser magnitude than that observed with Ca2+ currents in alpha1B-expressing cells. The subunits beta2 and gamma2 alone produced no effects, nor did constitutively active alpha s, alpha i1, and alpha o subunits. Phorbol esters enhanced alpha1E Ca2+ currents but had no effect on alpha1B currents, suggesting that protein kinase C activation was not responsible for the observed effects. When alpha1E Ca2+ currents were expressed without their beta subunits, they exhibited prepulse facilitation. These results demonstrate that alpha1E Ca2+ currents are less susceptible to direct modulation by G proteins than alpha1B currents and illustrate the antagonistic interactions between Ca2+ channel beta subunits and G proteins.
Shekter
L R
LR
Department of Pharmacological and Physiological Sciences, The University of Chicago, Illinois 60637, USA.
Taussig
R
R
Gillard
S E
SE
Miller
R J
RJ
D015220:Calcium Channels / Q000502:physiology*; D002460:Cell Line; D019204:GTP-Binding Proteins / Q000502:physiology*; D006801:Humans; D015640:Ion Channel Gating; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D014162:Transfection
D015220:Calcium Channels; D011994:Recombinant Proteins; D019204:GTP-Binding Proteins