Receptor--ion channel interactions in Torpedo electric organ: influence of thiol group modification

The influence of sulfhydryl and disulfide reagents on nicotinic cholinergic receptors and ion channels and their interactions was investigated using specific probes for receptor and channel binding sites in electric organs from Torpedo californica. N-ethylmaleimide, a sulfhydryl alkylating agent, did not alter receptor or ion channel binding, or receptor-mediated ion channel binding activation or desensitization. Alteration of receptor--ion channel coupling produced by treatment with an organic sulfhydryl, dithiothreitol, could be accounted for on the basis of decreases in receptor affinity for agonists. These changes were reversed by exposure to an oxidizing agent. Following reduction by dithiothreitol, N-ethylmaleimide treatment produced large decreases in the extent and affinity of both receptor and channel binding. No evidence for a direct role of thiol groups in ion channel function was obtained.