Antibodies directed against a nonapeptide sequence of the gamma-aminobutyrate (GABA)/benzodiazepine receptor alpha-subunit. Detection of a distinct alpha-like subunit in pig cerebral cortex but not cerebellum.
A synthetic peptide, corresponding to amino acid residues 101-109 of the bovine gamma-aminobutyrate/benzodiazepine receptor alpha-subunit, was used to raise a polyclonal antiserum. The reactivity of this antiserum towards polypeptides of both bovine and pig receptor preparations was established by immunoprecipitation and immunoblotting. Anti-peptide antibodies recognized the alpha-subunit (51 kDa) of receptor prepared from pig cerebellum or cerebral cortex. However, a polypeptide of 57 kDa was additionally recognized in cortical, but not cerebellar, preparations. This alpha-like polypeptide appeared larger than the band of polypeptides labelled irreversibly with [3H]muscimol (beta-subunit, 55-57 kDa) and corresponds to a polypeptide detected only in cortex after silver-staining or irreversible labelling with [3H]flunitrazepam. These results support the idea that the distinct regional patterns of polypeptides labelled irreversibly with [3H]flunitrazepam reflect the existence of heterologous distributions of distinct alpha-like subunits.
Kirkness
E F
EF
Department of Biochemistry, University of Leeds, U.K.
Turner
A J
AJ
D000345:Affinity Labels; D000818:Animals; D001923:Brain Chemistry; D002531:Cerebellum / Q000032:analysis; D002540:Cerebral Cortex / Q000032:analysis; D011232:Chemical Precipitation; D005445:Flunitrazepam; D015151:Immunoblotting; D009118:Muscimol; D010455:Peptides / Q000276:immunology*; D011963:Receptors, GABA-A; D013552:Swine
D000345:Affinity Labels; D010455:Peptides; D011963:Receptors, GABA-A; D009118:Muscimol; D005445:Flunitrazepam