Purification of the gamma-aminobutyric acidA/benzodiazepine receptor complex by immunoaffinity chromatography.
The bovine gamma-aminobutyric acidA/benzodiazepine receptor complex has been purified by a novel immunoaffinity chromatography method on immobilized monoclonal antibody 62-3G1. Immunopurification of the complex was achieved in a single step with an improved yield over affinity chromatography on the benzodiazepine Ro 7-1986/1. High-resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the immunoaffinity-purified receptor revealed three major peptide bands of 51,000, 55,000, and 57,000 Mr which were also present in the Ro 7-1986/1 affinity-purified receptor. Peptide mapping, immunoblotting with subunit specific antibodies, and photoaffinity labeling with [3H]flunitrazepam and [3H]muscimol have been used for the identification of receptor subunits, including several which comigrated in a single band in SDS-PAGE.
Park
D
D
Department of Neurobiology and Behavior, State University of New York, Stony Brook.
Vitorica
J
J
Tous
G
G
de Blas
A L
AL
D000345:Affinity Labels; D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D002540:Cerebral Cortex / Q000378:metabolism; D002846:Chromatography, Affinity; D007164:Immunosorbents; D008024:Ligands; D010449:Peptide Mapping; D010455:Peptides / Q000378:metabolism; D011963:Receptors, GABA-A / Q000302:isolation & purification* / Q000378:metabolism
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D007164:Immunosorbents; D008024:Ligands; D010455:Peptides; D011963:Receptors, GABA-A