Solution structures of chemoenzymatically synthesized heparin and its precursors

J Am Chem Soc. 2008 Oct 1;130(39):12998-3007. doi: 10.1021/ja8026345. Epub 2008 Sep 4.


We report the first chemoenzymatic synthesis of the stable isotope-enriched heparin from a uniformly labeled [(13)C,(15)N]N-acetylheparosan (-GlcA(1,4)GlcNAc-) prepared from E. coli K5. Glycosaminoglycan (GAG) precursors and heparin were formed from N-acetylheparosan by the following steps: chemical N-deacetylation and N-sulfonation leading to N-sulfoheparosan (-GlcA(1,4)GlcNS-); enzyme-catalyzed C5-epimerization and 2-O-sulfonation leading to undersulfated heparin (-IdoA2S(1,4)GlcNS-); enzymatic 6-O-sulfonation leading to the heparin backbone (-IdoA2S(1,4)GlcNS6S-); and selective enzymatic 3-O-sulfonation leading to the anticoagulant heparin, containing the GlcNS6S3S residue. Heteronuclear, multidimensional nuclear magnetic resonance spectroscopy was employed to analyze the chemical composition and solution structure of [(13)C,(15)N]N-acetylheparosan, precursors, and heparin. Isotopic enrichment was found to provide well-resolved (13)C spectra with the high sensitivity required for conformational studies of these biomolecules. Stable isotope-labeled heparin was indistinguishable from heparin derived from animal tissues and is a novel reagent for studying the interaction of heparin with proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Conformation
  • Carbohydrate Epimerases / chemistry
  • Carbohydrate Epimerases / metabolism
  • Carbon Isotopes
  • Glycosaminoglycans / chemical synthesis
  • Glycosaminoglycans / chemistry*
  • Heparin / chemical synthesis
  • Heparin / chemistry*
  • Heparitin Sulfate / chemical synthesis
  • Heparitin Sulfate / chemistry
  • Humans
  • Mice
  • Models, Molecular
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular
  • Oligosaccharides / chemical synthesis
  • Oligosaccharides / chemistry
  • Solutions
  • Sulfotransferases / chemistry
  • Sulfotransferases / metabolism


  • Carbon Isotopes
  • Glycosaminoglycans
  • N-acetylheparosan
  • Nitrogen Isotopes
  • Oligosaccharides
  • Solutions
  • Heparin
  • Heparitin Sulfate
  • Sulfotransferases
  • Carbohydrate Epimerases
  • heparin-heparan sulfate-glucuronic acid C5-epimerase