title: Influence of GABA(A) receptor α subunit isoforms on the benzodiazepine binding site.
abstract: Classical benzodiazepines, such as diazepam, interact with α(x)β(2)γ(2) GABA(A) receptors, x = 1, 2, 3, 5 and modulate their function. Modulation of different receptor isoforms probably results in selective behavioural effects as sedation and anxiolysis. Knowledge of differences in the structure of the binding pocket in different receptor isoforms is of interest for the generation of isoform-specific ligands. We studied here the interaction of the covalently reacting diazepam analogue 3-NCS with α(1)S204Cβ(2)γ(2), α(1)S205Cβ(2)γ(2) and α(1)T206Cβ(2)γ(2) and with receptors containing the homologous mutations in α(2)β(2)γ(2), α(3)β(2)γ(2), α(5)β(1/2)γ(2) and α(6)β(2)γ(2). The interaction was studied using radioactive ligand binding and at the functional level using electrophysiological techniques. Both strategies gave overlapping results. Our data allow conclusions about the relative apposition of α(1)S204Cβ(2)γ(2), α(1)S205Cβ(2)γ(2) and α(1)T206Cβ(2)γ(2) and homologous positions in α(2), α(3), α(5) and α(6) with C-atom adjacent to the keto-group in diazepam. Together with similar data on the C-atom carrying Cl in diazepam, they indicate that the architecture of the binding site for benzodiazepines differs in each GABA(A) receptor isoform α(1)β(2)γ(2), α(2)β(2)γ(2), α(3)β(2)γ(2), α(5)β(1/2)γ(2) and α(6)β(2)γ(2).
authors: Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.
authors: Benjamin P
authors: BP
authors: Lüscher
authors: Roland
authors: R
authors: Baur
authors: Maurice
authors: M
authors: Goeldner
authors: Erwin
authors: E
authors: Sigel
mesh_terms: D000595:Amino Acid Sequence; D001569:Benzodiazepines / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D003545:Cysteine / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D017354:Point Mutation; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011963:Receptors, GABA-A / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D020033:Protein Isoforms; D011963:Receptors, GABA-A; D001569:Benzodiazepines; D003545:Cysteine