title: Array-assisted characterization of a fucosyltransferase required for the biosynthesis of complex core modifications of nematode N-glycans.
abstract: Fucose is a common monosaccharide component of cell surfaces and is involved in many biological recognition events. Therefore, definition and exploitation of the specificity of the enzymes (fucosyltransferases) involved in fucosylation is a recurrent theme in modern glycosciences. Despite various studies, the specificities of many fucosyltransferases are still unknown, so new approaches are required to study these. The model nematode Caenorhabditis elegans expresses a wide range of fucosylated glycans, including N-linked oligosaccharides with unusual complex core modifications. Up to three fucose residues can be present on the standard N,N'-diacetylchitobiose unit of these N-glycans, but only the fucosyltransferases responsible for transfer of two of these (the core α1,3-fucosyltransferase FUT-1 and the core α1,6-fucosyltransferase FUT-8) were previously characterized. By use of a glycan library in both array and solution formats, we were able to reveal that FUT-6, another C. elegans α1,3-fucosyltransferase, modifies nematode glycan cores, specifically the distal N-acetylglucosamine residue; this result is in accordance with glycomic analysis of fut-6 mutant worms. This core-modifying activity of FUT-6 in vitro and in vivo is in addition to its previously determined ability to synthesize Lewis X epitopes in vitro. A larger scale synthesis of a nematode N-glycan core in vitro using all three fucosyltransferases was performed, and the nature of the glycosidic linkages was determined by NMR. FUT-6 is probably the first eukaryotic glycosyltransferase whose specificity has been redefined with the aid of glycan microarrays and so is a paradigm for the study of other unusual glycosidic linkages in model and parasitic organisms.
authors.affiliation: From the Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria and.
authors.forename: Shi
authors.initials: S
authors.lastname: Yan 闫石
authors.affiliation: the Biofunctional Nanomaterials Unit, CICbiomaGUNE, 20009 San Sebastian, Spain.
authors.forename: Sonia
authors.initials: S
authors.lastname: Serna
authors.affiliation: the Biofunctional Nanomaterials Unit, CICbiomaGUNE, 20009 San Sebastian, Spain.
authors.forename: Niels-Christian
authors.initials: NC
authors.lastname: Reichardt
authors.affiliation: From the Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria and.
authors.forename: Katharina
authors.initials: K
authors.lastname: Paschinger
authors.affiliation: From the Department für Chemie, Universität für Bodenkultur, A-1190 Wien, Austria and. Electronic address: iain.wilson@boku.ac.at.
authors.forename: Iain B H
authors.initials: IBH
authors.lastname: Wilson
mesh_terms: D000818:Animals; D053898:Biosynthetic Pathways; D017173:Caenorhabditis elegans / Q000201:enzymology*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D005643:Fucose / Q000737:chemistry / Q000378:metabolism; D005647:Fucosyltransferases / Q000378:metabolism*; D054794:Glycomics; D006031:Glycosylation; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D046228:Microarray Analysis; D050505:Mutant Proteins / Q000378:metabolism; D011134:Polysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D012996:Solutions; D013379:Substrate Specificity
chemical_list: D029742:Caenorhabditis elegans Proteins; D050505:Mutant Proteins; D011134:Polysaccharides; D012996:Solutions; D005643:Fucose; C584438:FUT-6 protein, C elegans; D005647:Fucosyltransferases
keywords: Glycan; Glycosyltransferases; Invertebrates; Mass Spectrometry (MS); Microarray; Oligosaccharide