title: Peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F cannot release glycans with fucose attached alpha 1----3 to the asparagine-linked N-acetylglucosamine residue.
abstract: The ability of peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F (PNGase F) from Flavobacterium meningosepticum and PNGase A from sweet almonds to deglycosylate N-glycopeptides and N-glycoproteins from plants was compared. Bromelain glycopeptide and horseradish peroxidase-C glycoprotein, which contain xylose linked beta 1----2 to beta-mannose and fucose linked alpha 1----3 to the innermost N-acetylglucosamine, were used as substrates. In contrast to PNGase A, the enzyme from F. meningosepticum did not act upon these substrates even at concentrations 100-fold higher than required for complete deglycosylation of commonly used standard substrates. After removal of alpha 1----3-linked fucose from the plant glycopeptide and glycoprotein by mild acid hydrolysis, they were readily degraded by PNGase F at moderate enzyme concentrations. Hence we conclude that alpha 1----3 fucosylation of the inner N-acetylglucosamine impedes the enzymatic action of PNGase F. Knowledge of this limitation of the deglycosylation potential of PNGase F may turn it from a pitfall into a useful experimental tool.
authors.affiliation: Institut für Chemie der Universität für Bodenkultur Wien, Austria.
authors.forename: V
authors.initials: V
authors.lastname: Tretter
authors.forename: F
authors.initials: F
authors.lastname: Altmann
authors.forename: L
authors.initials: L
authors.lastname: März
mesh_terms: D000117:Acetylglucosamine; D000581:Amidohydrolases / Q000378:metabolism*; D001216:Asparagine; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005417:Flavobacterium / Q000201:enzymology*; D005643:Fucose; D006020:Glycopeptides / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D013379:Substrate Specificity
chemical_list: D006020:Glycopeptides; D006023:Glycoproteins; D005643:Fucose; D001216:Asparagine; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D000117:Acetylglucosamine