Conversion of GalNAcβ(1–4)GlcNAcβ-OMe into GalNAcβ(1–4)-[Fucα(1–3)]GlcNAcβ-OMe using human milk α [formula omitted]-fucosyltransferase synthesis of a novel terminal element in glycoprotein glycans

Abstract

Incubation of GalNAcβ(1–4)GlcNAcβ-OMe with GDP-Fuc in the presence of human milk α 3 4 -fucosyltransferase resulted in the formation of GalNAcβ(1–4)[Fucα(1–3)]GlcNAcβ-OMe. Under conditions that led to complete α3-fucosylation of Galβ(1–4)GlcNAcβ-OEt, GalNAcβ(1–4)GlcNAcβ-OMe was fucosylated for more than 85%. For the identification of the isolated fucosylated products one- and two- dimensional 1H-NMR spectroscopy was applied. In vacuo molecular dynamics simulations of Galβ(1–4)[Fucα(1–3)]GlcNAcβ-OEt and GalNAcβ(1–4)[Fucα(1–3)]GlcNAcβ-OMe using the CHARMm based force field CHEAT, demonstrated only small differences between the conformations of these compounds. This illustrates the minor conformational influence of the substituent at C-2′, i.e. a hydroxyl function versus a N-acetyl group.