Sequencing Heparan Sulfate Saccharides

  • Jeremy E. Turnbull
Part of the Springer Protocols Handbooks book series (SPH)


The functions of the heparan sulfates (HSs) are determined by specific saccharide motifs within HS chains. These sequences confer selective protein binding properties and the ability to modulate protein activities (1,2). HS chains consist of an alternating disaccharide repeat of glucosamine (GlcN; N-acetylated or N-sulfated) and uronic acid (glucuronic [GlcA] or iduronic acid [IdoA]). The initial biosynthetic product containing N-acetylglucosamine (GlcNAc) and GlcA is modified by N-sulfation of the GlcN, ester (O)-sulfation (at positions 3 and 6 on the GlcN and position 2 on the uronic acids) and by epimerization of GlcA to IdoA. The extent of these modifications is incomplete and their degree and distribution varies in HS between different cell types. In HS chains N- and O-sulfated sugars are predominantly clustered in sequences of up to eight disaccharide units separated by N-acetyl-rich regions with relatively low sulfate content (3).


Uronic Acid Ammonium Persulfate Electrophoresis Buffer Sodium Cyanoborohydride Hexuronic Acid 
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  1. 1.
    Turnbull, J. E., Powell, A., and Guimond, S. E. (2001) Heparan sulphate: decoding a dynamic multifunctional cellular regulator. Trends Cell Biol. 11, 75–82.PubMedCrossRefGoogle Scholar
  2. 2.
    Bernfield, M., Gotte, M., Park, P. W., et al. (1999) Functions of cell surface heparan sulfate proteoglycans. Annu. Rev. Biochem. 68, 729–777.PubMedCrossRefGoogle Scholar
  3. 3.
    Turnbull, J. E. and Gallagher, J. T. (1991) Distribution of iduronate-2-sulfate residues in HS: evidence for an ordered polymeric structure. Biochem. J. 273, 553–559.PubMedGoogle Scholar
  4. 4.
    Turnbull, J. E., Fernig, D., Ke, Y., Wilkinson, M. C., and Gallagher, J. T. (1992) Identification of the basic FGF binding sequence in fibroblast HS. J. Biol. Chem. 267, 10,337–10,341.PubMedGoogle Scholar
  5. 5.
    Pervin, A., Gallo, C., Jandik, K., Han, X., and Linhardt, R. (1995) Preparation and structural characterisation of heparin-derived oligosaccharides. Glycobiology 5, 83–95.PubMedCrossRefGoogle Scholar
  6. 6.
    Yamada, S., Yamane, Y., Tsude, H., Yoshida, K., and Sugahara, K. (1998) A major common trisulfated hexasaccharide isolated from the low sulfated irregular region of porcine intestinal heparin. J. Biol. Chem. 273, 1863–1871.PubMedCrossRefGoogle Scholar
  7. 7.
    Yamada, S., Yoshida, K., Sugiura, M., Sugahara, K., Khoo, K., Morris, H., and Dell, A. (1993) Structural studies on the bacterial lyase-resistant tetrasaccharides derived from the antithrombin binding site of porcine mucosal intestinal heparin. J. Biol. Chem. 268, 4780–4787.PubMedGoogle Scholar
  8. 8.
    Mallis, L., Wang, H., Loganathan, D., and Linhardt, R. (1989) Sequence analysis of highly sulfated heparin-derived oligosaccharides using FAB-MS. Analyt. Chem. 61, 1453–1458.CrossRefGoogle Scholar
  9. 9.
    Rhomberg, A. J., Ernst, S., Sasisekharan, R., and Bieman, K. (1998) Mass spectrometric and capillary electrophoretic investigation of the enzymatic degradation of heparin-like glycosaminoglycans. Proc. Natl. Acad. Sci. USA 95, 4176–4181.PubMedCrossRefGoogle Scholar
  10. 10.
    Hopwood, J. (1989) Enzymes that degrade heparin and heparan sulfate, in Heparin (Lane and Lindahl, eds.), Edward Arnold Press, pp. 191–227.Google Scholar
  11. 11.
    Turnbull, J. E., Hopwood, J. J., and Gallagher, J. T. (1999) A strategy for rapid sequencing of heparan sulfate/heparin saccharides. Proc. Natl. Acad. Sci. USA 96, 2698–2703.PubMedCrossRefGoogle Scholar
  12. 12.
    Merry, C. L. R., Lyon, M., Deakin, J. A., Hopwood, J. J., and Gallagher, J. T. (1999) Highly sensitive sequencing of the sulfated domains of heparan sulfate. J. Biol. Chem. 274, 18,455–18,462.PubMedCrossRefGoogle Scholar
  13. 13.
    Vives, R. R., Pye, D. A., Samivirta, M., Hopwood, J. J., Lindahl, U., and Gallagher, J. T. (1999) Sequence analysis of heparan sulphate and heparin oligosaccharides. Biochem. J. 339, 767–773.PubMedCrossRefGoogle Scholar
  14. 14.
    Venkataraman, G., Shriver, Z., Ramar, R., and Sasisekharan, R. (1999) Sequencing complex poly saccharides. Science 286, 537–542.PubMedCrossRefGoogle Scholar
  15. 15.
    Guimond, S. E. and Turnbull, J. E. (1999) Fibroblast growth factor receptor signalling is dictated by specific heparan sulfate saccharides. Curr. Biol. 9, 1343–1346.PubMedCrossRefGoogle Scholar
  16. 16.
    Drummond, K. J., Yates, E. A., and Turnbull, J. (2001) Electrophoretic sequencing of heparin/heparan sulfate oligosaccharides using a highly sensitive fluorescent end label. Proteomics 1, 304–310.PubMedCrossRefGoogle Scholar
  17. 17.
    Shively, J. and Conrad, H. (1976) Formation of anhydrosugars in the chemical depolymerisation of heparin. Biochemistry 15, 3932–3942.PubMedCrossRefGoogle Scholar
  18. 18.
    Bienkowski and Conrad, H. (1985) Structural characterisation of the oligosaccharides formed by depolymerisation of heparin with nitrous acid. J. Biol. Chem. 260, 356–365.PubMedGoogle Scholar
  19. 19.
    Turnbull, J. E. and Gallagher, J. T. (1988) Oligosaccharide mapping of heparan sulfate by polyacrylamide-gradient-gel electrophoresis and electrotransfer to nylon membrane. Biochem. J. 251, 597–608.PubMedGoogle Scholar
  20. 20.
    Rice, K., Rottink, M., and Linhardt, R. (1987) Fractionation of heparin-derived oligosaccharides by gradient PAGE. Biochem. J. 244, 515–522.PubMedGoogle Scholar
  21. 21.
    Ludwigs, U., Elgavish, A., Esko, J., and Roden, L. (1987) Reaction of unsaturated uronic acid residues with mercuric salts. Biochem. J. 245, 795–804.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Jeremy E. Turnbull
    • 1
  1. 1.School of BiosciencesUniversity of BirminghamUK

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