Channelpedia

PubMed 12644584


Referenced in Channelpedia wiki pages of: none

Automatically associated channels: Cav2.2



Title: Mechanism of action of Gq to inhibit G beta gamma modulation of CaV2.2 calcium channels: probed by the use of receptor-G alpha tandems.

Authors: Federica Bertaso, Richard J Ward, Patricia Viard, Graeme Milligan, Annette C Dolphin

Journal, date & volume: Mol. Pharmacol., 2003 Apr , 63, 832-43

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/12644584


Abstract
The stable interaction of a G-protein coupled receptor and a particular partner G-protein was made possible by creating tandems between the alpha(2A) adrenergic receptor (alpha(2A)-R) and pertussis toxin-resistant mutants of different G alpha subunits of heterotrimeric G-proteins. Both alpha(2A)-R-G alpha(o) and alpha(2A)-R-G alpha(i) proved able to reconstitute agonist-induced voltage-dependent inhibition of N-type calcium channels (Ca(V)2.2) similar to the wild-type alpha(2A)-R when expressed in COS-7 cells. The interaction of G(q) with the G(i/o) signaling pathways was studied by expressing either G alpha(q) or a chimeric construct based on G alpha(q) containing the last five amino acids of G alpha(z), which is activated by alpha(2A)-R. It was found that G alpha(qz5) activated by the wild-type alpha(2A)-R inhibited Ca(V)2.2 currents in a voltage-independent fashion. Furthermore, G alpha(qz5) counteracted the voltage-dependent inhibition resulting from alpha(2A)-R-G alpha(o) activation. We subsequently investigated the basis for the behavior of G alpha(qz5). Our evidence suggests that this occurs as a result of a downstream effect of activation of G alpha(qz5) because it was blocked by C-terminal construct of phospholipase C beta 1. Furthermore it is likely to occur in part via protein kinase C (PKC) activation, because the PKC activator phorbol dibutyrate mimicked the effects of G alpha(qz5) in alpha(2A)-R-G alpha(o)-transfected cells. Conversely, cells expressing both alpha(2A)-R-G alpha(o) and G alpha(qz5) exhibited a partial restoration of voltage-dependent inhibition in the presence of the PKC inhibitor bisindolylmaleimide I (GF 109203X). The potential sites of phosphorylation are discussed.