Sequence analysis of heparan sulphate and heparin oligosaccharides

Romain R. Vivès, David A. Pye, Markku Salmivirta, John J. Hopwood, Ulf Lindahl, John T. Gallagher

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

The biological activity of heparan sulphate (HS) and heparin largely depends on internal oligosaccharide sequences that provide specific binding sites for an extensive range of proteins. Identification of such structures is crucial for the complete understanding of glycosaminoglycan (GAG)-protein interactions. We describe here a simple method of sequence analysis relying on the specific tagging of the sugar reducing end by 3H radiolabelling, the combination of chemical scission and specific enzymic digestion to generate intermediate fragments, and the analysis of the generated products by strong-anion-exchange HPLC. We present full sequence data on microgram quantities of four unknown oligosaccharides (three HS-derived hexasaccharides and one heparin-derived octasaccharide) which illustrate the utility and relative simplicity of the technique. The results clearly show that it is also possible to read sequences of inhomogeneous preparations. Application of this technique to biologically active oligosaccharides should accelerate progress in the understanding of HS and heparin structure-function relationships and provide new insights into the primary structure of these polysaccharides.

LanguageEnglish
Pages767-773
Number of pages7
JournalBiochemical Journal
Volume339
Issue number3
DOIs
Publication statusPublished - 1 May 1999

Keywords

  • Glycosaminoglycan
  • Lysosomal enzymes
  • Sequence

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Vivès, R. R., Pye, D. A., Salmivirta, M., Hopwood, J. J., Lindahl, U., & Gallagher, J. T. (1999). Sequence analysis of heparan sulphate and heparin oligosaccharides. Biochemical Journal, 339(3), 767-773. https://doi.org/10.1042/0264-6021:3390767
Vivès, Romain R. ; Pye, David A. ; Salmivirta, Markku ; Hopwood, John J. ; Lindahl, Ulf ; Gallagher, John T. / Sequence analysis of heparan sulphate and heparin oligosaccharides. In: Biochemical Journal. 1999 ; Vol. 339, No. 3. pp. 767-773.
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Vivès, RR, Pye, DA, Salmivirta, M, Hopwood, JJ, Lindahl, U & Gallagher, JT 1999, 'Sequence analysis of heparan sulphate and heparin oligosaccharides', Biochemical Journal, vol. 339, no. 3, pp. 767-773. https://doi.org/10.1042/0264-6021:3390767

Sequence analysis of heparan sulphate and heparin oligosaccharides. / Vivès, Romain R.; Pye, David A.; Salmivirta, Markku; Hopwood, John J.; Lindahl, Ulf; Gallagher, John T.

In: Biochemical Journal, Vol. 339, No. 3, 01.05.1999, p. 767-773.

Research output: Contribution to journalArticle

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Vivès RR, Pye DA, Salmivirta M, Hopwood JJ, Lindahl U, Gallagher JT. Sequence analysis of heparan sulphate and heparin oligosaccharides. Biochemical Journal. 1999 May 1;339(3):767-773. https://doi.org/10.1042/0264-6021:3390767