The effects of drugs on the incorporation of a conformationally-sensitive, hydrophobic probe into the ion channel of the nicotinic acetylcholine receptor.

The pattern of incorporation of the hydrophobic photolabel 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine([125I]TID) into the nicotinic acetylcholine receptor (AChR) is a sensitive measure of AChR conformation (resting state or desensitized). We determined the ability of tetracaine, dibucaine, procaine, lidocaine, chlorpromazine or phencyclidine to inhibit [125I]TID photolabeling of the AChR as a function of drug concentration, both as a measure of the ability of these drugs to desensitize the AChR, and to characterize the [125I]TID binding site. To localize the site(s) of drug action, experiments were performed in the absence and presence of saturating concentrations of alpha-bungarotoxin (BgTx), to block drug binding to the agonist binding site. On the basis of the concentration dependence of their effects, which was not altered by the presence of BgTx, tetracaine and dibucaine appeared to block [125I]TID incorporation competitively, suggesting that the high-affinity [125I]TID binding site is the non-competitive blocker binding site presumed to exist in the interior of the AChR ion channel. Procaine, chlorpromazine, lidocaine and phencyclidine blocked [125I]TID incorporation at lower concentrations in the absence of BgTx than in its presence, suggesting that these drugs block incorporation by inducing desensitization when bound to their high-affinity non-competitive blocker binding sites and that BgTx countered the drug effect by allosterically stabilizing the resting state.