Article date: 1986/3/1
PubMed ID: 2419499
Journal name: Journal of neurochemistry (ISSN: 0022-3042)
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.
Author List: Stephenson F A, Casalotti S O, Mamalaki C, Barnard E A
Publication Types: Journal Article; Research Support, Non-U.S. Gov't
Substances mentioned in the article: Antibodies; Antigens; Bridged Bicyclo Compounds; Bridged Bicyclo Compounds, Heterocyclic; Carbolines; Epitopes; Receptors, GABA-A; Muscimol; Flunitrazepam; tert-butylbicyclophosphorothionate; propyl beta-carboline-3-carboxylate;
Mesh terms: Allosteric Regulation; Animals; Antibodies/immunology; Antibody Specificity; Antigens/immunology; Binding Sites; Bridged Bicyclo Compounds/metabolism; Bridged Bicyclo Compounds, Heterocyclic; Carbolines/metabolism; Cattle; Cerebral Cortex/analysis; Electrophoresis, Polyacrylamide Gel; Epitopes/immunology; Flunitrazepam/metabolism; Immunologic Techniques; Immunosorbent Techniques; Muscimol/metabolism; Receptors, GABA-A/immunology;