GDP-fucose: beta-N-acetylglucosamine (Fuc to (Fuc alpha 1----6GlcNAc)-Asn-peptide)alpha 1----3-fucosyltransferase activity in honeybee (Apis mellifica) venom glands. The difucosylation of asparagine-bound N-acetylglucosamine.
Incubation of honeybee (Apis mellifica) venom-gland extracts with GDP-[14C]fucose and GlcNAc beta 1----2Man alpha 1----6(GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4(Fuc alpha 1----6)GlcNAc beta 1----N-Asn-peptide(NAc) gave a labeled product in 40% yield. Analysis by 500-MHz 1H-NMR spectroscopy indicated the transferred fucose-(Fuc) residue to be alpha 1----3-linked to the Asn-bound GlcNAc. Further proof was provided by one-dimensional and two-dimensional 1H-NMR analysis of the incubation mixture, after incubation with beta-N-acetylhexosaminidase. The established carbohydrate structure (formula; see text) proves the existence of a novel alpha 1----3-fucosyltransferase with the ability to effect difucosylation of the Asn-bound GlcNAc in N-glycans.
Staudacher
E
E
Institut für Chemie, Universität für Bodenkultur, Vienna, Austria.
Altmann
F
F
Glössl
J
J
März
L
L
Schachter
H
H
Kamerling
J P
JP
Hård
K
K
Vliegenthart
J F
JF
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001514:Bee Venoms; D001516:Bees / Q000201:enzymology*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005647:Fucosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D006154:Guanosine Diphosphate Fucose / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D013379:Substrate Specificity
D001514:Bee Venoms; D006154:Guanosine Diphosphate Fucose; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase; D000117:Acetylglucosamine