Conversion of GalNAc beta(1-4)GlcNAc beta-OMe into GalNAc beta (1-4)-[Fuc alpha(1-3)]GlcNAc beta-OMe using human milk alpha 3/4-fucosyltransferase. Synthesis of a novel terminal element in glycoprotein glycans.
Incubation of GalNAc beta(1-4)GlcNAc beta-OMe with GDP-Fuc in the presence of human milk alpha 3/4-fucosyltransferase resulted in the formation of GalNAc beta(1-4)[Fuc alpha(1-3)]GlcNAc beta-OMe. Under conditions that led to complete alpha 3-fucosylation of Gal beta(1-4)GlcNAc beta-OEt, GalNAc beta(1-4)GlcNAc beta-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 Gal beta(1-4)[Fuc alpha(1-3)]GlcNAc beta-OEt and GalNAc beta(1-4)[Fuc alpha(1-3)]GlcNAc beta-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.
Bergwerff
A A
AA
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
van Kuik
J A
JA
Schiphorst
W E
WE
Koeleman
C A
CA
van den Eijnden
D H
DH
Kamerling
J P
JP
Vliegenthart
J F
JF
D002240:Carbohydrate Sequence; D005647:Fucosyltransferases / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D016256:Lewis X Antigen / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008895:Milk, Human / Q000201:enzymology*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism
D006023:Glycoproteins; D016256:Lewis X Antigen; D009844:Oligosaccharides; D011134:Polysaccharides; D005647:Fucosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase