Cross-talk between G-protein and protein kinase C modulation of N-type calcium channels is dependent on the G-protein beta subunit isoform.
The modulation of N-type calcium current by protein kinases and G-proteins is a factor in the fine tuning of neurotransmitter release. We have previously shown that phosphorylation of threonine 422 in the alpha(1B) calcium channel domain I-II linker region resulted in a dramatic reduction in somatostatin receptor-mediated G-protein inhibition of the channels and that the I-II linker consequently serves as an integration center for cross-talk between protein kinase C (PKC) and G-proteins (Hamid, J., Nelson, D., Spaetgens, R., Dubel, S. J., Snutch, T. P., and Zamponi, G. W. (1999) J. Biol. Chem. 274, 6195-6202). Here we show that opioid receptor-mediated inhibition of N-type channels is affected to a lesser extent compared with that seen with somatostatin receptors, hinting at the possibility that PKC/G-protein cross-talk might be dependent on the G-protein subtype. To address this issue, we have examined the effects of four different types of G-protein beta subunits on both wild type and mutant alpha(1B) calcium channels in which residue 422 has been replaced by glutamate to mimic PKC-dependent phosphorylation and on channels that have been directly phosphorylated by protein kinase C. Our data show that phosphorylation or mutation of residue 422 antagonizes the effect of Gbeta(1) on channel activity, whereas Gbeta(2), Gbeta(3), and Gbeta(4) are not affected. Our data therefore suggest that the observed cross-talk between G-proteins and protein kinase C modulation of N-type channels is a selective feature of the Gbeta(1) subunit.
Cooper
C B
CB
Departments of Physiology & Biophysics and Pharmacology & Therapeutics, Neuroscience and Smooth Muscle Research Groups, University of Calgary, Calgary, Alberta T2N 4N1, Canada.
Arnot
M I
MI
Feng
Z P
ZP
Jarvis
S E
SE
Hamid
J
J
Zamponi
G W
GW
D020864:Calcium Channels, N-Type / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D010766:Phosphorylation; D020033:Protein Isoforms / Q000502:physiology; D011493:Protein Kinase C / Q000502:physiology*; D021122:Protein Subunits; D009435:Synaptic Transmission
D020864:Calcium Channels, N-Type; D020033:Protein Isoforms; D021122:Protein Subunits; D011493:Protein Kinase C; D019204:GTP-Binding Proteins