Solution Structures of Chemoenzymatically Synthesized Heparin and Its Precursors

  1. Zhang, Z. 2
  2. McCallum, S.A. 2
  3. Xie, J. 2
  4. Nieto, L. 1
  5. Corzana, F. 4
  6. Jiménez-Barbero, J. 1
  7. Chen, M. 3
  8. Liu, J. 3
  9. Linhardt, R.J. 2
  1. 1 Centro de Investigaciones Biológicas
    info

    Centro de Investigaciones Biológicas

    Madrid, España

    ROR https://ror.org/04advdf21

  2. 2 Rensselaer Polytechnic Institute
    info

    Rensselaer Polytechnic Institute

    Troy, Estados Unidos

    ROR https://ror.org/01rtyzb94

  3. 3 University of North Carolina System
    info

    University of North Carolina System

    Chapel Hill, Estados Unidos

    ROR https://ror.org/0566a8c54

  4. 4 Universidad de La Rioja
    info

    Universidad de La Rioja

    Logroño, España

    ROR https://ror.org/0553yr311

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Revista:
Journal of the American Chemical Society

ISSN: 0002-7863

Año de publicación: 2008

Volumen: 130

Número: 39

Páginas: 12998-13007

Tipo: Artículo

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DOI: 10.1021/JA8026345 PMID: 18767845 SCOPUS: 2-s2.0-58149158096 WoS: WOS:000259553700044 GOOGLE SCHOLAR

Otras publicaciones en: Journal of the American Chemical Society

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Resumen

We report the first chemoenzymatic synthesis of the stable isotope-enriched heparin from a uniformly labeled [13C,15N]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 [13C,15N]N-acetylheparosan, precursors, and heparin. Isotopic enrichment was found to provide well-resolved 13C 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. © 2008 American Chemical Society.