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Use of the “core-2”-N-acetylglucosaminyltransferase in the chemical-enzymatic synthesis of a sialyl-Lex-containing hexasaccharide found on O-linked glycoproteins

Authors
Journal
Carbohydrate Research
0008-6215
Publisher
Elsevier
Publication Date
Volume
244
Issue
1
Identifiers
DOI: 10.1016/0008-6215(93)80011-3
Disciplines
  • Biology

Abstract

Abstract A simple preparation of the “core-II” N-acetylglucosaminyltransferase (UDP- d-Glc pNAc:β- d-Gal p-(1 → 3)-α- d-Gal pNAc (GlcNAc to GalNAc) β-(1 → 6)-GlcNAc-transferase, GlcNAcT, EC 2.4.1.102) from commercial mouse kidney acetone powder is reported. The enzyme obtained in a single step of affinity chromatography is suitable for use in preparative oligosaccharide synthesis. In conjunction with previously described preparations of β-(1 → 4)-galactosyltransferase (EC 2.4.1.22), α-(2 → 3)-sialyltransferase (EC 2.4.99.6) and α-(1 → 3/4)-fucosyltransferase (EC 2.4.1.65), the GlcNAcT was used in the first step of a sequence which converted the disaccharide β- d-Gal p-(1 → 3)-α- d-Gal pNAc-OR into the sialyl-Le x-containing structure α- d-Neu pAc-(2 → 3)-β- d-Gal p-(1 → 4)-[α- l-Fuc p-(1 → 3)]-β- d-Glc pNAc-(1 → 6)-β- d-Gal p-(1 → 3)-α- d-Gal pNAc-OR ( 5), where R = CH 2) 8CO 2Me. Hexasaccharide 5, thus assembled in only one week once the enzymes were prepared, was characterized by 1H and 13C NMR spectroscopy and fast-atom bombardment mass spectrometry, as were all intermediate oligosaccharides. The core II GlcNAcT thus joins the expanding repertoire of readily available reagents for the rapid assembly of oligosaccharides.

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