Conversion of GalNAcβ(1–4)GlcNAcβ‐OMe into GalNAcβ(1–4)‐[Fucα(1–3)]GlcNAcβ‐OMe using human milk α3/4‐fucosyltransferase synthesis of a novel terminal element in glycoprotein glycans

FEBS Letters

van Kuik, J.Albert²  Schiphorst, Wietske E.C.M.¹  Vliegenthart, Johannes F.G.²  van den Eijnden, Dirk H.¹  Koeleman, Carolien A.M.¹  Bergwerff, Aldert A.²  Kamerling, Johannis P.² 

[1] Department of Medical Chemistry, Vrije Universiteit, Amsterdam, The Netherlands;Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, PO Box 80075, 3508 TB Utrecht, The Netherlands

关键词: Fucosyltransferase;    GalNAc-containing N-glycan;    Conformational analysis;    Lewis x;    Fuc;    fucose;    Neu5Ac;    N-acetylneuraminic acid;    Lex;    Lewis x;    ID;    one dimensional;    2D;    two-dimensional;    COSY;    correlation spectroscopy;    HOHAHA;    homonuclear Hartmann—Hahn;    MLEV;    composite pulse devised by Malcom Levitt;    r.O.e.;    rotating frame nuclear Overhauser enhancement;    ROESY;    rotating frame nuclear Overhauser enhancement spectroscopy;    CHEAT;    carbohydrate hydroxyls represented by extended atoms;    CHARMm;    Molecular Simulations Inc. version of Chemistry Harvard macromolecular mechanics;    MD;    molecular dynamics;   

DOI  :  10.1016/0014-5793(93)81698-Y

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Incubation of GalNAcβ(1–4)GlcNAcβ-OMe with GDP-Fuc in the presence of human milk α3/4-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.

【 授权许可】

Unknown