Enzymatic synthesis of the alpha 3-galactosyl-Lex tetrasaccharide: a potential ligand for selectin-type adhesion molecules.
Using recombinant UDP-Gal:Gal beta 1-->4GlcNAc alpha 1,3-galactosyltransferase and human milk alpha 1,3-fucosyltransferase the disaccharide Gal beta 1-->4GlcNAc has been converted in vitro into a tetrasaccharide product. The product has been characterized by gel filtration chromatography and HPLC and was analyzed using 1H-NMR. Based on NMR spectral data along with the known linkage specificity of the alpha 1,3-galactosyltransferase and the alpha 1,3-fucosyltransferase used, the chromatographic behaviour of the product, and the 1:1 molar ratios of the galactose and fucose residues calculated from incorporated radioactivity, it is concluded that the structure of the tetrasaccharide product is Gal alpha 1-->3Gal beta 1--4[Fuc alpha 1-->3]-GlcNAc. The tetrasaccharide is a non-charged analogue of the sialyl-Lex determinant that potentially may act as a ligand structure in selectin-mediated cell-cell adhesion.
Joziasse
D H
DH
Department of Medical Chemistry, Vrije Universiteit, Amsterdam, The Netherlands.
Schiphorst
W E
WE
Koeleman
C A
CA
Van den Eijnden
D H
DH
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005647:Fucosyltransferases / Q000378:metabolism*; D005700:Galactosyltransferases / Q000378:metabolism*; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000096:biosynthesis*; D009682:Magnetic Resonance Spectroscopy; D008895:Milk, Human / Q000201:enzymology; D008969:Molecular Sequence Data; D009036:Moths; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism
D007983:Lewis Blood Group Antigens; D009844:Oligosaccharides; D011994:Recombinant Proteins; D005647:Fucosyltransferases; D005700:Galactosyltransferases; C031692:galactoside 3-fucosyltransferase; C037959:beta-D-galactosyl-N-acetylglucosaminylglycopeptide alpha-1,3-galactosyltransferase