title: Glycosyltransferases in chemo-enzymatic synthesis of oligosaccharides.
abstract: Many oligosaccharides are not commercially available, which limits studies focused on elucidation of glycan functions; therefore chemo-enzymatic methods to synthesize them can be very useful. Here, we describe the procedure to synthesize the Galα1-3GalNAcβ1-4GlcNAcβ-R (Gal-LDN) moiety, containing the Galα1-3GalNAc epitope found on the parasitic helminth Haemonchus contortus. An acceptor substrate providing a terminal N-acetylglucosamine was prepared by coupling the fluorescent hydrophobic aglycon, 2,6-diaminopyridine (DAP), to N,N'-diacetylchitobiose. By the subsequent action of recombinant Caenorhabditis elegans β1,4-N-acetylgalactosaminyltransferase the substrate was efficiently converted to GalNAcβ1-4GlcNAcβ-R (LDN-R). Since no recombinant α1,3-galactosyltransferase has been described that acts on terminal βGalNAc, we used bovine α1,3-galactosyltransferase to obtain a partial conversion of LDN-R to the Gal-LDN antigen. This method can be applied to synthesize any oligosaccharide, provided that specific glycosyltransferases are available, or related enzymes that can be pushed to elongate the selected acceptor.
authors.affiliation: Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Boris
authors.initials: B
authors.lastname: Tefsen
authors.forename: Irma
authors.initials: I
authors.lastname: van Die
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D017173:Caenorhabditis elegans / Q000201:enzymology; D002240:Carbohydrate Sequence; D002417:Cattle; D004187:Disaccharides / Q000737:chemistry; D000939:Epitopes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D016695:Glycosyltransferases / Q000378:metabolism*; D006188:Haemonchiasis / Q000469:parasitology; D006189:Haemonchus / Q000737:chemistry* / Q000276:immunology; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D011725:Pyridines / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000947:Antigens, Helminth; D004187:Disaccharides; D000939:Epitopes; D009844:Oligosaccharides; D011725:Pyridines; D011994:Recombinant Proteins; C032438:chitobiose; D016695:Glycosyltransferases; C505797:2,6-diaminopyridine