title: Host cell-specific folding and assembly of the neuronal nicotinic acetylcholine receptor alpha7 subunit.
abstract: Expression of the cloned neuronal nicotinic acetylcholine receptor (nAChR) alpha7 subunit in several cultured mammalian cell lines has revealed that the folding, assembly, and subcellular localization of this protein are critically dependent upon the nature of the host cell. In all cell lines that were examined, high levels of alpha7 protein were detected by metabolic labelling and immunoprecipitation after transfection with the cloned alpha7 cDNA. In contrast, elevated levels of alpha-bungarotoxin binding could be detected in only two of the nine cell lines. Both of these "alpha7-permissive" cell lines [rat phaeochromocytoma (PC12) and human neuroblastoma (SH-SY5Y)] express an endogenous alpha7 subunit. However, by expression of an epitope-tagged alpha7 subunit, it has been possible to show that the elevation in surface alpha-bungarotoxin binding in these two cell lines is due to expression of cDNA-encoded alpha7. The cell-specific misfolding of the neuronal nAChR alpha7 subunit is a phenomenon that is not shared by either the hetero-oligomeric muscle nAChR or the homooligomeric serotonin receptor 5-HT3 subunit. Our data also indicate that the cell-specific misfolding cannot be explained by a requirement for the coassembly with other known nAChR subunits and cannot be alleviated by treatments that have been reported to affect the assembly efficiency of other neurotransmitter-gated ion channels.
authors.affiliation: Department of Pharmacology, University College London, England.
authors.forename: S T
authors.initials: ST
authors.lastname: Cooper
authors.forename: N S
authors.initials: NS
authors.lastname: Millar
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002038:Bungarotoxins / Q000378:metabolism; D002460:Cell Line; D002645:Chickens; D000242:Cyclic AMP / Q000378:metabolism; D018076:DNA, Complementary; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D008968:Molecular Conformation; D009132:Muscles / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000502:physiology; D016716:PC12 Cells; D017510:Protein Folding; D051381:Rats; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D018076:DNA, Complementary; D011978:Receptors, Nicotinic; D000242:Cyclic AMP