title: Role of N-linked oligosaccharides in the biosynthetic processing of the cystic fibrosis membrane conductance regulator.
abstract: The epithelial chloride channel CFTR is a glycoprotein that is modified by two N-linked oligosaccharides. The most common mutant CFTR protein in patients with cystic fibrosis, DeltaF508, is misfolded and retained by ER quality control. As oligosaccharide moieties of glycoproteins are known to mediate interactions with ER lectin chaperones, we investigated the role of N-linked glycosylation in the processing of wild-type and DeltaF508 CFTR. We found that N-glycosylation and ER lectin interactions are not major determinants of trafficking of wild-type and DeltaF508 from the ER to the plasma membrane. Unglycosylated CFTR, generated by removal of glycosylation sites or treatment of cells with the N-glycosylation inhibitor tunicamycin, did not bind calnexin, but did traffic to the cell surface and exhibited chloride channel activity. Most importantly, unglycosylated DeltaF508 CFTR still could not escape quality control in the early secretory pathway and remained associated with the ER. However, the absence of N-linked oligosaccharides did reduce the stability of wild-type CFTR, causing significantly more-rapid turnover in post-ER compartments. Surprisingly, the individual N-linked carbohydrates do not play equivalent roles and modulate the fate of the wild-type protein in different ways in its early biosynthetic pathway.
authors.affiliation: Mayo Clinic College of Medicine, Mayo Clinic Arizona, Scottsdale, AZ 85259, USA.
authors.forename: Xiu-Bao
authors.initials: XB
authors.lastname: Chang
authors.forename: April
authors.initials: A
authors.lastname: Mengos
authors.forename: Yue-Xian
authors.initials: YX
authors.lastname: Hou
authors.forename: Liying
authors.initials: L
authors.lastname: Cui
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Jensen
authors.forename: Andrei
authors.initials: A
authors.lastname: Aleksandrov
authors.forename: John R
authors.initials: JR
authors.lastname: Riordan
authors.forename: Martina
authors.initials: M
authors.lastname: Gentzsch
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D037281:Calnexin / Q000378:metabolism; D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008954:Models, Biological; D018832:Molecular Chaperones / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D050505:Mutant Proteins / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D021381:Protein Transport
chemical_list: D037102:Lectins; D018832:Molecular Chaperones; D050505:Mutant Proteins; D009844:Oligosaccharides; C093349:cystic fibrosis transmembrane conductance regulator delta F508; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D037281:Calnexin