Peptide heterogeneity of GABAA/benzodiazepine receptors in bovine cerebral cortex and cerebellum.
The GABAA/benzodiazepine receptor complex has been purified from both bovine cerebral cortex and cerebellum by immunoaffinity chromatography on immobilized monoclonal antibody 62-3G1. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified receptor from either cerebral cortex or cerebellum revealed 3 main bands corresponding to 51,000, 55,000 and 57,000 Mr silver-stained peptides. In addition, a minor band corresponding to a 53,000 Mr peptide was also found. The differences between the two receptor preparations were: (1) that the main silver-stained 55,000 Mr subunit was present in a relative smaller quantity in cerebellum than in cerebral cortex, and (2) when the membrane-bound receptor was photoaffinity-labeled with [3H]flunitrazepam and subsequently immunoaffinity-purified, two photolabeled peptide bands of 51,000 and 57,000 Mr were found in cerebral cortex while only the 51,000 Mr photolabeled peptide was detected in cerebellum following one-dimension sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Peptide maps of the 57,000 Mr [3H]flunitrazepam photoaffinity-labeled peptide indicated that it was composed of two closely migrating photolabeled peptides of 55,000 Mr and 57,000 Mr. Peptide mapping and deglycosylation experiments using the [3H]flunitrazepam photolabeled receptor suggested that the photolabeled peptides commonly present in cerebellum and cerebral cortex are qualitatively similar if not identical. The results suggest that there are subunits of some type(s) of GABAA R/BZDR complex(es) which are more abundant in cerebral cortex than in cerebellum. Photoaffinity labeling with [3H]muscimol showed similar photolabeled peptides in both cerebral cortex and cerebellum: two main peptides of 54,000 and 57,000 Mr were photolabeled with [3H]muscimol to a similar extent in both receptor preparations.(ABSTRACT TRUNCATED AT 250 WORDS)
Park
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Department of Neurobiology and Behavior, State University of New York, Stony Brook.
de Blas
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D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002531:Cerebellum / Q000378:metabolism*; D002540:Cerebral Cortex / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005445:Flunitrazepam / Q000378:metabolism; D008970:Molecular Weight; D009118:Muscimol / Q000378:metabolism; D009928:Organ Specificity; D010449:Peptide Mapping; D011963:Receptors, GABA-A / Q000302:isolation & purification* / Q000378:metabolism
D011963:Receptors, GABA-A; D009118:Muscimol; D005445:Flunitrazepam