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The GABAA receptor family in the mammalian brain

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Abstract

The GABAA-benzodiazepine receptor protein from bovine brain was purified by affinity chromatography and the subunit composition examined by gel electrophoresis in sodium dodecyl sulfate. Protein staining revealed a doublet at 51–53 kDa, a band at 55 kDa, and a broad band at 57–59 kDa. The 51 and 53 kDa bands co-migrated with the α1 and α2 gene products identified by Western blotting with subtype-specific antibodies. These two bands were also photoaffinity labeled by [3H]flunitrazepam, as was a breakdown product at 44 kDa. Partial sequencing of proteolytic fragments of these polypeptides yielded sequences found in all α clones, and identified the benzodiazepine binding site within residues 8–297 and probably between 106–297 of α1; the 44 kDa and 31 kDa bands yielded fragments containing α3 sequence. The native α3 polypeptide was identified with subtype-specific antibody at 57 kDa overlapping with the two major bands photolabeled with [3H]muscimol at 55 and 58 kDa. Antisera to a β-selective peptide recognized four bands at 60, 58, 57 and 55 kDa. Thus, one can identify 6–8 distinct polypeptides with the possibility of another 4–6 in purified GABAA receptor proteins, depending on brain region, consistent with the family of gene products suggested by molecular cloning.

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  1. Michel H. Bureau

    Present address: Laboratoire de Biochimie Générale et Comparee, 21 Place Delcour, 4020, Liege, Belgium

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  1. Department of Pharmacology, School of Medicine, Brain Research Institute, and Mental Retardation Research Center, University of California, 90024, Los Angeles, CA

    Richard W. Olsen, Michel H. Bureau, Shuichi Endo & Geoffrey Smith

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Olsen, R.W., Bureau, M.H., Endo, S. et al. The GABAA receptor family in the mammalian brain. Neurochem Res 16, 317–325 (1991). https://doi.org/10.1007/BF00966095

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