m-Sulfonate benzene diazonium chloride: a powerful affinity label for the gamma-aminobutyric acid binding site from rat brain.
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.
Bouchet
M J
MJ
Laborátoire de Chimie Bio-Organique, URA 1386 du CNRS, Faculté de Pharmacie, Université Louis Pasteur Strasbourg, Illkirch, France.
Jacques
P
P
Ilien
B
B
Goeldner
M
M
Hirth
C
C
D000345:Affinity Labels; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D003979:Diazonium Compounds; D008024:Ligands; D009118:Muscimol / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013425:Sulfanilic Acids; D013574:Synaptosomes / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D000345:Affinity Labels; D003979:Diazonium Compounds; D008024:Ligands; D013425:Sulfanilic Acids; D009118:Muscimol; D005680:gamma-Aminobutyric Acid; C000469:diazobenzenesulfonic acid