Interaction of p-aminophenyldichloroarsine, an arsenical with specificity for vicinal cysteines, with [3H]cytisine binding sites in rat brain membranes.

The arsenical compound p-aminophenyldichloroarsine (APA) is selective for spatially close thiols with which it forms a stable complex. The alpha subunits of nicotinic acetylcholine receptors are defined by the presence of a pair of adjacent cysteines close to the agonist binding site. Here the interaction of APA with [3H]cytisine binding sites, which correspond to the major subtype of nicotinic receptors in rat brain has been examined. Incubation of brain membranes with 10 microM APA abolished [3H]cytisine binding. The action of APA was dependent on prior reduction of sulphydryls with dithiothreitol. APA effects could not be reversed by oxidizing agents but could be reversed by the antiarsenical reagent 2,3-dimercapto-1-propane sulphonic acid. Under the conditions used, the concentration of APA producing a half-maximal decrease in binding was 130 nM. The loss of [3H]cytisine binding was due to a decrease in the number of binding sites (Bmax) with no effect on affinity for the radioligand (Kd). Nicotinic ligands failed to protect against the reduction and arsenylation of neuronal receptor sites. These observations are consistent with the potent interaction of APA with this neuronal nicotinic receptor.