title: Preparation and characterization of (15)N-enriched, size-defined heparan sulfate precursor oligosaccharides.
abstract: We report the preparation of size-defined [(15)N]N-acetylheparosan oligosaccharides from Escherichia coli-derived (15)N-enriched N-acetylheparosan. Optimized growth conditions of E. coli in minimal media containing (15)NH(4)Cl yielded [(15)N]N-acetylheparosan on a preparative scale. Depolymerization of [(15)N]N-acetylheparosan by heparitinase I yielded resolvable, even-numbered oligosaccharides ranging from disaccharide to icosaccharide. Anion-exchange chromatography-assisted fractionation afforded size-defined [(15)N]N-acetylheparosan oligosaccharides identifiable by ESI-TOFMS. These isotopically labeled oligosaccharides will prove to be valuable research tools for the chemoenzymatic synthesis of heparin and heparan sulfate oligosaccharides and for the study of their structural biology.
authors: Interdepartmental Program in Neuroscience, University of Utah, Salt Lake City, 20 North 1900 East, MREB #401, UT 84132, USA.
authors: Crystal
authors: C
authors: Sigulinsky
authors: Ponnusamy
authors: P
authors: Babu
authors: Xylophone V
authors: XV
authors: Victor
authors: Balagurunathan
authors: B
authors: Kuberan
mesh_terms: D006493:Heparin / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry*; D009587:Nitrogen Isotopes / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013329:Structure-Activity Relationship
chemical_list: D006493:Heparin; D006497:Heparan Sulfate; D009587:Nitrogen Isotopes; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins