Quantitative immunoprecipitation studies with anti-gamma-aminobutyric acidA receptor gamma 2 1-15 Cys antibodies.
Antibodies raised against the synthetic peptide NH2-QKSDDDYEDYASNKTC-COOH (gamma 2 1-15 Cys), which corresponds to the N-terminal amino acid sequence with a C-terminal cysteine of the human gamma 2 subunit of the gamma-aminobutyric acidA (GABAA) receptor, were used to study the quantitative immunoprecipitation of agonist benzodiazepine binding sites from bovine brain. Anti-gamma 2 1-15 Cys antibodies were found to immunoprecipitate specifically in parallel [3H]flunitrazepam- and [3H]muscimol-reversible binding sites in a dose-dependent manner. The maximum percentages of [3H]flunitrazepam binding sites immunoprecipitated from detergent extracts of bovine cerebral cortex, cerebellum, and hippocampus were 68, 77, and 83%, respectively. Immunoprecipitation studies with anti-alpha 1 324-341 antibodies carried out in parallel with anti-gamma 2 1-15 Cys antibodies provided evidence for the promiscuity of the gamma 2 subunit within native GABAA receptors. These results substantiate the association of the gamma 2 polypeptide with native GABAA receptors.
Duggan
M J
MJ
Department of Pharmaceutical Chemistry, School of Pharmacy, University of London, England.
Pollard
S
S
Stephenson
F A
FA
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002417:Cattle; D005445:Flunitrazepam / Q000378:metabolism; D008969:Molecular Sequence Data; D009118:Muscimol / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology*; D011233:Precipitin Tests / Q000379:methods*; D011963:Receptors, GABA-A / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D014018:Tissue Distribution
D000906:Antibodies; D010446:Peptide Fragments; D011963:Receptors, GABA-A; D009118:Muscimol; D005445:Flunitrazepam