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...

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Bibliographic Details
Published in:Carbohydrate research 2010-01, Vol.345 (2), p.250-256
Main Authors: Sigulinsky, Crystal, Babu, Ponnusamy, Victor, Xylophone V, Kuberan, Balagurunathan
Format: Article
Language:English
Subjects:
Heparin - chemistry
Heparitin Sulfate - chemistry
Nitrogen Isotopes - chemistry
Oligosaccharides - chemistry
Receptors, Cell Surface - chemistry
Receptors, Cell Surface - metabolism
Spectrometry, Mass, Electrospray Ionization
Structure-Activity Relationship
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Summary: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.
ISSN:1873-426X
DOI:10.1016/j.carres.2009.10.024