Preparation and characterization of (15)N-enriched, size-defined heparan sulfate precursor oligosaccharides.
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.
Sigulinsky
Crystal
C
Interdepartmental Program in Neuroscience, University of Utah, Salt Lake City, 20 North 1900 East, MREB #401, UT 84132, USA.
Babu
Ponnusamy
P
Victor
Xylophone V
XV
Kuberan
Balagurunathan
B
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
D006493:Heparin; D006497:Heparan Sulfate; D009587:Nitrogen Isotopes; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins