Antibodies recognising the GABAA/benzodiazepine receptor including its regulatory sites.
Polyclonal antibodies have been raised against the GABA/benzodiazepine receptor purified to homogeneity from bovine cerebral cortex in deoxycholate and Triton X-100 media. Radioimmunoassay was applied to measure specific antibody production using the 125I-labelled gamma-aminobutyric acid (GABA)/benzodiazepine receptor as antigen. The antibodies specifically immunoprecipitated the binding sites for [3H]muscimol and for [3H]flunitrazepam from purified preparations. In addition, when a 3-[(3-cholamidopropyl)dimethylammonio] 1-propanesulphonate (CHAPS) extract of bovine brain membranes was treated with the antibodies, those sites as well as the [3H]propyl-beta-carboline-3-carboxylate binding, the [35S]t-butylbicyclophosphorothionate binding (TBPS), the barbiturate-enhanced [3H]flunitrazepam binding, and the GABA-enhanced [3H]flunitrazepam binding were all removed together into the immunoprecipitate. Western blot experiments showed that these antibodies recognise the alpha-subunit of the purified GABA/benzodiazepine receptor. These results further support the existence in the brain of a single protein, the GABAA receptor, containing a set of regulatory binding sites for benzodiazepines and chloride channel modulators.
F A
FA
Stephenson
S O
SO
Casalotti
C
C
Mamalaki
E A
EA
Barnard
D000494:Allosteric Regulation; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D001665:Binding Sites; D001643:Bridged Bicyclo Compounds / Q000378:metabolism; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002243:Carbolines / Q000378:metabolism; D002417:Cattle; D002540:Cerebral Cortex / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005445:Flunitrazepam / Q000378:metabolism; D007158:Immunologic Techniques; D007163:Immunosorbent Techniques; D009118:Muscimol / Q000378:metabolism; D011963:Receptors, GABA-A / Q000276:immunology* / Q000378:metabolism
D000906:Antibodies; D000941:Antigens; D001643:Bridged Bicyclo Compounds; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002243:Carbolines; D000939:Epitopes; D011963:Receptors, GABA-A; D009118:Muscimol; D005445:Flunitrazepam; C037476:tert-butylbicyclophosphorothionate; C028585:propyl beta-carboline-3-carboxylate