Evidence for the Existence of Differential O‐Glycosylated α5‐Subunits of the γ‐Aminobutyric AcidA Receptor in the Rat Brain
- 1 January 1993
- journal article
- Published by Wiley in Journal of Neurochemistry
- Vol. 60 (1) , 93-98
- https://doi.org/10.1111/j.1471-4159.1993.tb05826.x
Abstract
Polyclonal antibodies were raised to synthetic peptides having amino acid sequences corresponding with the N‐ or C‐terminal part of the γ‐aminobutyric acidA (GABAA) receptor α5‐subunit. These anti‐peptide α5(2–10) or anti‐peptide α5(427–433) antibodies reacted specifically with GABAA receptors purified from the brains of 5–10‐day‐old rats in an enzyme‐linked immunosorbent assay and were able to dose‐dependently immunoprecipitate up to 6.3 or 13.1% of the GABAA receptors present in the incubation, respectively. In immunoblots, each of these antibodies reacted with the same two protein bands with apparent molecular mass of 53 or 57 kDa. After exhaustive treatment of purified GABAA receptors with N‐Glycanase, each of these antibodies identified two proteins with apparent molecular masses of 46 and 48 kDa. Additional treatment of GABAA receptors with neuraminidase and O‐Glycanase resulted in an apparently single protein with molecular mass of 47 kDa, which again was identified by both the anti‐peptide α5(2–10) and the anti‐peptide α5(427–433) antibody. These results indicate the existence of at least two different α5‐sub‐units of the GABAA receptor that differ in their carbohydrate content. In contrast to other α‐ or β‐subunits of GABAA receptors so far investigated, at least one of these two α5‐subunits contains O‐linked carbohydrates.Keywords
This publication has 26 references indexed in Scilit:
- GABAA receptor subtypes immunopurified from rat brain with α subunit-specific antibodies have unique pharmacological propertiesNeuron, 1991
- Cloning, pharmacological characteristics and expression pattern of the rat GABAA receptor α4 subunitFEBS Letters, 1991
- Identification of α1-, α2- and α3-subunit isoforms of the GABAA-benzodiazepine receptor in the rat brainNeuroscience Letters, 1991
- N‐Deglycosylation and immunological identification indicates the existence of β‐subunit isoforms of the rat GABAA receptorFEBS Letters, 1991
- γ‐Aminobutyric AcidA Receptor Heterogeneity Is Increased by Alternative Splicing of a Novel β‐Subunit Gene TranscriptJournal of Neurochemistry, 1991
- Generation of Two Forms of the γ‐Aminobutyric AcidA Receptor γ‐2‐Subunit in Mice by Alternative SplicingJournal of Neurochemistry, 1991
- Function and pharmacology of multiple GABAA receptor subunitsTrends in Pharmacological Sciences, 1991
- Identification of α2‐ and α3‐subunits of the GABAA‐benzodiazepine receptor complex purified from the brains of young ratsFEBS Letters, 1990
- Functional expression and sites of gene transcription of a novel α subunit of the GABAA receptor in rat brainFEBS Letters, 1990
- Evidence for the existence of several different α- and β-subunits of the GABA/benzodiazepine receptor complex from rat brainNeuroscience Letters, 1989