TITLE:
The action of ethanol on G protein. In silico and cellular/molecular evidences
AUTHORS:
Pamela Fernandez, Jessica Moreno, Claudio Barrientos, Sergio A. Aguila, Daniela Leon, Sebastián Ortiz, Ramon Silva, Francisco Rodriguez, Maritza Leonardi, Violeta Morin, Ximena Romo
KEYWORDS:
Alcoholism; Ethanol; Glycine Receptor; G Proteins; Signals Transduction
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.4 No.5,
May
30,
2013
ABSTRACT: Ethanol
(EtOH) enhances glycinergic currents in the central nervous system (CNS).
Because evidence for an interaction between the α1 subunit of the glycine receptor (α1GlyR) and the G protein Gβγ subunit exists in vitro and because cAMP levels are known
to increase in response to EtOH, we wanted to investigate the interaction
between Gβγ and α1GlyR in response to EtOH
treatment in HEK293 cells and to explore the possible sites of interaction
between EtOH and the Gαs
subunit. His pull-down assays in GlyR-His6-transfected HEK293 cells incubated
with ethanol or propofol revealed that only EtOH treatment increased the
binding of Gβγ heterodimers to α1GlyR. Using molecular modelling
(protein structure prediction), was modelled the hGαs protein for the first time and
validated this model by site-directed mutagenesis. By molecular docking, we
identified some potential regions of interaction between hGαs and EtOH that are located on the SIII and SI regions of the Gαs. Therefore, we conclude that
ethanol increases the interaction between α1GlyR and Gβγ in HEK293 cells, an effect that might be attributed to the interaction between
EtOH and hGαs, which consequently
stimulates hGαs.