title: Identification of the glycosylation site of the adenovirus type 5 fiber protein.
abstract: The fiber protein purified from the pool of nonincorporated viral protein after infection of cells with adenovirus type 5 exists as two forms separable by reverse-phase HPLC. As determined by mass spectrometry, this heterogeneity results from a difference in one O-linked N-acetylglucosamine (GlcNac). A western blot analysis using a monoclonal antibody directed against the GlcNac motif showed that only one of the two forms reacted with the antibody, suggesting that one form carries a single GlcNac and the other form has none. The ratio of glycosylated to nonglycosylated forms of fiber, which is about 1, is conserved in assembled viruses. After digestion of glycosylated fiber with endoproteinase GluC, isolation of the glycosylated peptide by reverse-phase HPLC, and chemical derivatization using dimethylamine, the site of glycosylation was located in the fiber shaft at serine 109 by mass spectrometry. Elimination of glycosylation by site-directed mutagenesis of fiber should help to understand the function of this postranslational modification.
authors.affiliation: Transgene SA, 11 rue de Molsheim, 67082 Strasbourg, France. gilles.cauet@biopep.com
authors.forename: Gilles
authors.initials: G
authors.lastname: Cauet
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Strub
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Leize
authors.forename: Elsa
authors.initials: E
authors.lastname: Wagner
authors.forename: Alain
authors.initials: A
authors.lastname: Van Dorsselaer
authors.forename: Monika
authors.initials: M
authors.lastname: Lusky
mesh_terms: D000256:Adenoviridae / Q000737:chemistry*; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D008969:Molecular Sequence Data; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D014764:Viral Proteins