Assembly and N-glycosylation of all ACh receptor subunits are required for their efficient insertion into plasma membranes.

Various combinations of synthetic acetylcholine receptor (AChR) subunit mRNAs were injected into Xenopus oocytes, and assembly of incomplete AChRs and their insertion into the plasma membrane was studied. Assembly of incomplete AChRs is not greatly affected by the absence of one or two of the other subunits. In contrast, the membrane insertion of incomplete AChRs is profoundly reduced as compared with complete AChRs. The role of N-glycosylation on the assembly of AChR subunits, and on their insertion into plasma membranes, was also studied by using the Xenopus oocyte expression system and tunicamycin. Assembly of non-N-glycosylated AChR subunits occurs in tunicamycin-treated oocytes, but these subunits remain in intracellular compartments, suggesting that N-glycosylation of AChR subunits is not a prerequisite for receptor assembly, but is required for their efficient insertion into the plasma membrane.