Subunit selectivity and epitope characterization of mAbs directed against the GABAA/benzodiazepine receptor

• Published in: The Journal of cell biology Vol. 110; no. 6; pp. 2043 - 2048
• Main Authors: EWERT, M, SHIVERS, B. D, LUÊDDENS, H, MOÊHLER, H, SEEBURG, P. H
• Format: Journal Article
• Language: English
• Published: New York, NY Rockefeller University Press 01.06.1990; The Rockefeller University Press
• Subjects: Amino Acid Sequence; Antibodies, Monoclonal - immunology; Base Sequence; Biological and medical sciences; Cell Line, Transformed; Cell receptors; Cell structures and functions; Epitopes - immunology; Fundamental and applied biological sciences. Psychology; Humans; Immunohistochemistry; Kidney - cytology; Kidney - embryology; Kidney - ultrastructure; Molecular and cellular biology; Molecular Sequence Data; Neuropeptide receptors; Receptors, GABA-A - analysis; Receptors, GABA-A - immunology; Receptors, GABA-A - metabolism; Characterization; Human; Gabaergic receptor; Cell culture; Transfection; Antigenic determinant; Immunocytochemistry; Aminoacid; GABA; Neurotransmitter; Monoclonal antibody; Kidney
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.110.6.2043

mAbs bd 17, bd 24, and bd 28 raised against bovine cerebral gamma-aminobutyric acid (GABAA)/benzodiazepine receptors were analyzed for their ability to detect each of 12 GABAA receptor subunits expressed in cultured mammalian cells. Results showed that mAb bd 17 recognizes epitopes on both beta 2 and beta 3 subunits while mAb bd 24 is selective for the alpha 1 subunit of human and bovine, but not of rat origin. The latter antibody reacts with the rat alpha 1 subunit carrying an engineered Leu at position four, documenting the first epitope mapping of a GABAA receptor subunit-specific mAb. In contrast to mAbs bd 17 and bd 24, mAb bd 28 reacts with all GABAA receptor subunits tested but not with a glycine receptor subunit, suggesting the presence of shared epitopes on subunits of GABA-gated chloride channels.