Skip to main content
SpringerLink
Log in

A single step method for purification of sulfated oligosaccharides

  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

Purifying and analysing sulfated oligosaccharides by mass spectrometry often constitutes a challenge. We present here a single step method to isolate sulfated compounds from a complex mixture of neutral and acidic oligosaccharide–alditols. The strategy relies on the exclusion of sulfated molecules from strong cation exchange resin. By testing a wide range of reduced mucin type O-glycans isolated from different biological sources, we demonstrate that this method permits, without prior chemical modification, the specific purification of sulfate-containing oligosaccharides present in any quantity from very complex mixtures of molecules.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Abbreviations

dHex:

deoxyHexose

DHB:

2,5-dihydroxybenzoic acid

ESI:

ElectroSpray Ionization

HPLC:

High Performance Liquid Chromatography

Kdn:

2-keto-3-deoxynononic acid

MALDI-TOF:

Matrix Assisted Laser Desorption Ionisation-Time of Flight

MS:

Mass Spectrometry

Man:

Mannose

S:

Sulfate group

TLC:

Thin Layer Chromatography

References

  1. Degroote, S., Ducourouble, M.P., Roussel, P., Lamblin, G.: Sequential biosynthesis of sulfated and/or sialylated Lewis x determinants by transferases of the human bronchial mucosa. Glycobiology 9, 1199–1211 (1999)

    Article  PubMed  CAS  Google Scholar 

  2. Degroote, S., Maes, E., Humbert, P., Delmotte, P., Lamblin, G., Roussel, P.: Sulfated oligosaccharides isolated from the respiratory mucins of a secretor patient suffering from chronic bronchitis. Biochimie 85, 369–379 (2003)

    Article  PubMed  CAS  Google Scholar 

  3. Lamblin, G., Degroote, S., Perini, J-M., Delmotte, P., Scharfman, A., Davril, M., Lo-Guidice, J-M., Houdret, N., Dumur, V., Klein, A., Roussel, P.: Human airway mucin glycosylation: a combinatory of carbohydrate determinants which vary in cystic fibrosis. Glycoconj. J. 18, 661–684 (2001)

    Article  PubMed  CAS  Google Scholar 

  4. Spiro, R.G., Bhoyroo, V.D.: Occurrence of sulfate in the asparagine-linked complex carbohydrate units of thyroglobulin-identification and localization of galactose-3-sulfate and N-acetylglucosamine-6-sulfate residues in the human and calf proteins. J. Biol. Chem. 263, 14351–14358 (1988)

    PubMed  CAS  Google Scholar 

  5. Hooper, L.V., Beranek, M.C., Manzella, S.M., Baenziger, J.U.: Differential expression of GaINAc-4-sulfotransferase and GaINAc-transferase results in distinct glycoforms of carbonic anhydrase VI in parotid and submaxillary glands. J. Biol. Chem. 270, 5985–5993 (1995)

    Article  PubMed  CAS  Google Scholar 

  6. Hård, K., Van Zadelhoff, C., Moonen, P., Kamerhing, J.P., Vliegenthart, J.F.C.: The Asn-linked carbohydrate chains of human Tamm-Horssfal glycoprotein of one male-novel sulfated and novel N-acetylgalactosamine containing N-linked carbohydrate chains. Eur. J. Biochem. 209, 895–915 (1992)

    Article  PubMed  Google Scholar 

  7. Yuen, C.-T., Lawson, A.M., Chai, W., Larkin, M., Stoll, M.S., Stuart, A.C., Sullivan, F.X., Ahem, T.J., Feizi, T.: Novel sulfated ligands for the cell adhesion molecule E-selectin revealed by the neoglycolipid technology among O-linked ohigosaccharides on an ovarian cystadenoma glycoprotein. Biochemistry 31, 9126–9131 (1992)

    Article  PubMed  CAS  Google Scholar 

  8. Burger, D., Simon, M., Perruisseau, G., Steck, A.J.: The epitope(s) recognized by HNK-1 antibody and IgM paraprotein in neuropathy is present on several N-linked oligosaccharide structures on human P0 and myelin-associated glycoprotein. J. Neurochem. 54, 1569–1575 (1990)

    Article  PubMed  CAS  Google Scholar 

  9. Fiete, D., Srivastava, V., Hindsgaul, O., Baenziger, J.U.: A hepatic reticuloendothelial cell receptor specific for S04–4GalNAcb1,4GlcNAcb that mediates rapid clearance of lutropin. Cell 67, 1103–1110 (1991)

    Article  PubMed  CAS  Google Scholar 

  10. Hemmerich, S., Leffler, H., Rosen, S.D.: Structure of the O-glycans in GlyCAM-1, an endothelial-derived ligand for L-selectin. J. Biol. Chem. 270, 12035–12047 (1995)

    Article  PubMed  CAS  Google Scholar 

  11. Thomsson, K.A., Schulz, B.L., Packer, N.H., Karlsson, N.G.: MUC5B glycosylation in human saliva reflects blood group and secretor status. Glycobiology 15, 791–804 (2005)

    Article  PubMed  CAS  Google Scholar 

  12. Lo-Guidice, J.M., Wieruszeski, J.M., Lemoine, J., Verbert, A., Roussel, P., Lamblin, G.: Sialylation and sulfation of the carbohydrate chains in respiratory mucins from a patient with cystic fibrosis. J. Biol. Chem. 269, 18794–18813 (1994)

    PubMed  CAS  Google Scholar 

  13. Robbe, C., Capon, C., Maes, E., Rousset, M., Zweibaum, A., Zanetta, J.P., Michalski, J.C.: Evidence of regio-specific glycosylation in human intestinal mucins: presence of an acidic gradient along the intestinal tract. J. Biol. Chem. 278, 46337–46348 (2003)

    Article  PubMed  CAS  Google Scholar 

  14. Thornton, D.J., Sheehan, J.K.: From mucins to mucus: toward a more coherent understanding of this essential barrier. Proc. Am. Thorac. Soc. 1, 54–61 (2004)

    Article  PubMed  CAS  Google Scholar 

  15. Davril, M., Degroote, S., Humbert, P., Galabert, C., Dumur, V., Lafitte, J.J., Lamblin, G., Roussel, P.: The sialylation of bronchial mucins secreted by patients suffering from cystic fibrosis or from chronic bronchitis is related to the severity of airway infection. Glycobiology 9, 311–321 (1999)

    Article  PubMed  CAS  Google Scholar 

  16. Scharfman, A., Delmotte, P., Beau, J., Lamblin, G., Roussel, P., Mazurier, J.: Sialyl-Le(x) and sulfo-sialyl-Le(x) determinants are receptors for P. aeruginosa. Glycoconj J. 17, 735–740 (2000)

    Article  PubMed  CAS  Google Scholar 

  17. Scharfman, A., Arora, S.K., Delmotte, P., Van Brussel, E., Mazurier, J., Ramphal, R., Roussel, P.: Recognition of Lewis x derivatives present on mucins by flagellar components of Pseudomonas aeruginosa. Infect. Immun. 69, 5243–5248 (2001)

    Article  PubMed  CAS  Google Scholar 

  18. Karlsson, N.G., Karlsson, H., Hansson, G.C.: Strategy for the investigation of O-linked oligosaccharides from mucins based on the separation into neutral, sialic acid- and sulfate-containing species. Glycoconj. J. 12, 69–76 (1995)

    Article  PubMed  CAS  Google Scholar 

  19. Ciucanu, I., Kerek, F.: A simple and rapid method for the permethylation of carbohydrates. Carbohyd. Res. 131, 209–217 (1984)

    Article  CAS  Google Scholar 

  20. Maes, E., Florea, D., Delplace, F., Lemoine, J., Plancke, Y., Strecker, G.: Structural analysis of the oligosaccharide–alditols released by reductive b-elimination from oviducal mucins of Rana temporaria. Glycoconj. J. 14, 127–146 (1997)

    Article  PubMed  CAS  Google Scholar 

  21. Florea, D., Maes, E., Strecker, G.: Primary structure of seven sulfated oligosaccharide–alditols released by reductive b-elimination from oviducal mucins of Rana temporaria. Characterization of the sequence HSO3(3)GlcA(b1–3)Gal. Carbohydr. Res. 302, 179–189 (1997)

    Article  PubMed  CAS  Google Scholar 

  22. Coppin, A., Maes, E., Morelle, W., Strecker, G.: Structural analysis of 13 neutral oligosaccharide–alditols released by reductive b-elimination from oviducal mucins of Rana temporaria. Eur J Biochem. 266, 94–104 (1999)

    Article  PubMed  CAS  Google Scholar 

  23. Strecker, G., Wieruszeski, J.M., Michalski, J.C., Alonso, C., Boilly, B., Montreuil, J.: Characterization of Le(x), Le(y) and A Le(y) antigen determinants in KDN-containing O-linked glycan chains from Pleurodeles waltl jelly coat eggs. FEBS Lett. 298, 39–43 (1992)

    Article  PubMed  CAS  Google Scholar 

  24. Strecker, G., Wieruszeski, J.M., Michalski, J.C., Montreuil, J.: 1H-and 13C-n.m.r. spectroscopy of 2-oxo-3-deoxy-d-glycero-d-galactononulosonic acid-containing oligosaccharide–alditols bearing Lewis X, Lewis Y and A-Lewis Y determinants isolated from the jelly coat of Pleurodeles waltl eggs. Biochem. J. 287, 905–909 (1992)

    PubMed  CAS  Google Scholar 

  25. Kamerling, J.P., Vliegenthart, J.F.G.: High-resolution 1H-nuclear magnetic resonance spectroscopy of oligosaccharide–alditols released from mucin-type O-glycoproteins. Biol. Magn. Res. 10, 1–288 (1992)

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Unité de Glycobiologie Structurale et Fonctionnelle, CNRS UMR 8576, USTL, 59655, Villeneuve d’Ascq, France

    Estelle Garenaux, Doina Florea, Gérard Strecker & Yann Guérardel

  2. Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei, 11529, Taiwan

    Shin-Yi Yu & Kay-Hooi Khoo

  3. Institute of Biochemical Sciences, National Taiwan University, Taipei, 106, Taiwan

    Shin-Yi Yu & Kay-Hooi Khoo

Authors
  1. Estelle Garenaux
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shin-Yi Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Doina Florea
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gérard Strecker
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kay-Hooi Khoo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yann Guérardel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yann Guérardel.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM

ESM 1 (PDF 363 KB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Garenaux, E., Yu, SY., Florea, D. et al. A single step method for purification of sulfated oligosaccharides. Glycoconj J 25, 903–915 (2008). https://doi.org/10.1007/s10719-008-9139-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10719-008-9139-3

Keywords

Search

Navigation