Background: Propofol is a sedative agent that at clinical concentrations acts
by allosterically activating or potentiating the γ-aminobutyric acid type A (GABAA)
receptor. Mutational, modeling, and photolabeling studies with propofol and its
analogues have identified potential interaction sites in the transmembrane domain of
the receptor. At the "+" of the β subunit, in the β-α interface, meta-azipropofol labels
the M286 residue in the third transmembrane domain. Substitution of this residue
with tryptophan results in loss of potentiation by propofol. At the "-" side of the β
subunit, in the α-β interface (or β-β interface, in the case of homomeric β receptors),
ortho-propofol diazirine labels the H267 residue in the second transmembrane
domain. Structural modeling indicates that the β(H267) residue lines a cavity that
docks propofol with favorable interaction energy.
Method: We used two-electrode voltage clamp to determine the functional effects of mutations to the "+"
and "-" sides of the β subunit on activation of the α1β3 GABAA receptor by propofol.
Results: We found that while the individual mutations had a small effect, the combination of the M286W
mutation with tryptophan mutations of selected residues at the α-β interface leads to strong reduction in
gating efficacy for propofol.
Conclusion: We conclude that α1β3 GABAA receptors can be activated by propofol interactions with the
β-β, α-β, and β-α interfaces, where distinct, non-equivalent regions control channel gating. Any interface
can mediate activation, hence substitutions at all interfaces are required for loss of activation by propofol.