E. Sigel et al., Role of the conserved lysine residue in the middle of the predicted extracellular loop between M2 and M3 in the GABA(A) receptor, J NEUROCHEM, 73(4), 1999, pp. 1758-1764
In alpha 1, beta 2, and gamma 2 subunits of the gamma-aminobutyric acid A (
GABA(A)) receptor, a conserved lysine residue occupies the position in the
middle of the predicted extracellular loop between the transmembrane M2 and
M3 regions. In all three subunits, this residue was mutated to alanine, Wh
ereas the mutation in alpha 1 and beta 2 subunits resulted each in about a
sixfold shift of the concentration-response curve for GABA to higher concen
trations, no significant effect by mutation in the gamma subunit was detect
ed. The affinity for the competitive inhibitor bicuculline methiodide was n
ot affected by the mutations in either the alpha 1 subunit or the beta 2 su
bunit. Concentration-response curves for channel activation by pentobarbita
l were also shifted to higher concentrations by the mutation in the alpha a
nd beta subunits. Binding of [H-3]Ro 15-1788 was unaffected by the mutation
in the alpha subunit, whereas the binding of [H-3]muscimol was shifted to
lower affinity. Mutation of the residue in the alpha 1 subunit to E, Q, or
R resulted in an about eight-, 10-, or fivefold shift, respectively, to hig
her concentrations of the concentration-response curve for GABA. From these
observations, it is concluded that the corresponding residues on the al an
d beta 2 subunits are involved more likely in the gating of the channel by
GABA than in the binding of GABA or benzodiazepines.