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Nicotinic acetylcholine receptor was localized in a receptor-rich membrane preparation from the electric organ of Torpedo californica by applying an immunoferritin technique. The membrane preparation was incubated with (Fab')2 fragments derived from specific rabbit antibodies against the purified acetylcholine receptor and subsequently with ferritin-conjugated goat antiserum to rabbit immunoglobulin. More than 50% of the vesicles were found to be labeled with ferritin while the rest remained unlabeled. Ferritin labeling on both sides of the membrane was evident in open membrane vesicles, whereas in closed vescles the labeling was confined to the outer surface due to the inability of the tracer to penetrate the membrane. These data suggest that antigenic sites of the receptor molecule are exposed on both sides of the excitable membrane, and that acetylcholine receptor may be a transmembrane protein.

proceedings of the national academy of sciences of the united states of america

Localization of acetylcholine receptor in excitable membrane from the electric organ of Torpedo: Evidence for exposure of receptor antigenic sites on both sides of the membrane.