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

• Published in: FEBS letters Vol. 334; no. 1; pp. 133 - 138
• Main Authors: Bergwerff, Aldert A., van Kuik, J.Albert, Schiphorst, Wietske E.C.M., Koeleman, Carolien A.M., van den Eijnden, Dirk H., Kamerling, Johannis P., Vliegenthart, Johannes F.G.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 1993
• Subjects: Conformational analysis; Fucosyltransferase; GalNAc-containing N-glycan; Lewis x; Le x, Lewis x; MLEV, composite pulse devised by Malcom Levitt; Conformational analysis; ID, one dimensional; Fucosyltransferase; r.O.e., rotating frame nuclear Overhauser enhancement; COSY, correlation spectroscopy; Fuc, fucose; CHEAT, carbohydrate hydroxyls represented by extended atoms; CHARMm, Molecular Simulations Inc. version of Chemistry Harvard macromolecular mechanics; MD, molecular dynamics; Lewis x; ROESY, rotating frame nuclear Overhauser enhancement spectroscopy; 2D, two-dimensional; Neu5Ac, N-acetylneuraminic acid; GalNAc-containing N-glycan; HOHAHA, homonuclear Hartmann—Hahn
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(93)81698-Y

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.