Article date: 1992/10/1
PubMed ID: 1402891
Journal name: Journal of neurochemistry (ISSN: 0022-3042)
m-Sulfonate benzene diazonium chloride (MSBD) was used to affinity-label the gamma-aminobutyric acid (GABA) binding site from rat brain membranes. To assess the irreversibility of the labeling reaction, we used an efficient ligand dissociation procedure combined to a rapid [3H]muscimol binding assay, both steps being performed on filter-adsorbed membranes. Inactivation of specific [3H]-muscimol binding sites by MSBD and its prevention by GABA were both time- and concentration-dependent. The time course of MSBD labeling was shortened as the pH of the incubation medium was increased from 6.2 to 8. These data suggest that MSBD can efficiently label the GABA binding site through alkylation of a residue having an apparent dissociation constant around neutrality.
Author List: Bouchet M J, Jacques P, Ilien B, Goeldner M, Hirth C
Publication Types: Journal Article; Research Support, Non-U.S. Gov't
Substances mentioned in the article: Affinity Labels; Diazonium Compounds; Ligands; Sulfanilic Acids; Muscimol; gamma-Aminobutyric Acid; diazobenzenesulfonic acid;
Mesh terms: Affinity Labels; Animals; Binding Sites; Brain/metabolism; Diazonium Compounds; Ligands; Muscimol/metabolism; Rats; Rats, Wistar; Sulfanilic Acids; Synaptosomes/metabolism; gamma-Aminobutyric Acid/metabolism;