Purification of Gastrointestinal Mucins and Analysis of Their O-Linked Oligosaccharides

  • Barry J. Campbell
  • Jonathan M. Rhodes
Part of the Methods in Molecular Biology™ book series (MIMB, volume 76)


The role of mucus in the protection of intestinal, bronchial, nasopharyngeal, and cervical mucosae has long been recognized, but poorly understood. Research has been stimulated by the expectation that an underlying mucus abnormality might be present, not only in diseases such as cystic fibrosis where there is an obvious physical alteration in mucus, but also in conditions such as inflammatory bowel disease, peptic ulceration, and intestinal cancer where mucus abnormalities at present seem more subtle (1). Progress has been relatively slow, however, not least because of the difficulties in obtaining a pure but undamaged mucus glycoprotein (mucin) preparation.


Sialic Acid HPLC Grade Water Fast Protein Liquid Chromatography Pulse Amperometric Detection Benzamidine Hydrochloride 
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  1. 1.
    Rhodes, J. M., Campbell, B. J., and Finnie, I. A. (1994) Mucus and the gastrointestinal tract, in Recent Advances in Gastroenterology 10 (Pounder, R., ed.), Churchill-Livingstone, Edinburgh, pp 57–79.Google Scholar
  2. 2.
    Finnie, I. A., Dwarakanath, A. D., Taylor, B. A., and Rhodes, J. M. (1995) Colonic mucin synthesis is increased by sodium butyrate. Gut 36, 93–99.CrossRefGoogle Scholar
  3. 3.
    Dwarakanath, A. D., Campbell, B. J., Tsai, H. H., Sunderland, D., Hart, C. A., and Rhodes, J. M. (1995) Fecal mucinase activity assessed in inflammatory bowel disease using 14C-threonine labelled mucin substrate. Gut 37, 58–62.CrossRefGoogle Scholar
  4. 4.
    Sheehan, J. K. and Carlstedt, I. (1987) Size heterogeniety of cervical mucus glycoproteins-studies performed with rate-zonal centrifugation and laser light scattering. Biochem. J. 245, 757–762.Google Scholar
  5. 5.
    Campbell, B. J., Finnie, I. A., Hounsell, E. F., and Rhodes, J. M. (1995) Direct demonstration of increased expression of Thomsen-Friedenreich (TF) antigen from colonic adenocarcinoma and ulcerative colitis mucin and its concealment in normal mucin. J. Clin. Invest. 95, 571–576.CrossRefGoogle Scholar
  6. 6.
    Makin, C. A. (1986) Monoclonal antibodies raised to colorectal carcinoma antigens. Ann. R. Coll. Surg. Eng. 68, 298–301Google Scholar
  7. 7.
    Endo, Y. and Kobata, A. (1976) Partial purification and characterisation of an endo-α-N-acetylgalactosaminidase from the culture medium of Diplococcus pneumoniae J. Biochem. 80, 1–8.Google Scholar
  8. 8.
    Umemoto, J., Matta, K. L., Barlow, J. J., and Bhavanandan, V. P. (1978) Action of endo-α-N-acetylgalactosaminidase on synthetic glycosidases including chromogenic substrates. Anal. Biochem. 91, 186–193.CrossRefGoogle Scholar
  9. 9.
    Parker, N., Raouf, A. H., Finnie, I. A., Ryder, S. D., Campbell, B. J., Tsai, H. H., Iddon, D., Milton, J. D., and Rhodes, J. M. (1993) High performance gel filtration using monodisperse highly cross-linked agarose as a one step system for mucin purification. Blamed Chromatogr. 7, 68–74.CrossRefGoogle Scholar
  10. 10.
    Lo-Guidice, J. M., Wieruszeski, J. M., Lemome, J., Verbert, A., Roussel, P., and Lamblin, G. (1994) Sialylation and sulphation of the carbohydrate chains in respiratory mucins from a patient with cystic fibrosis. J. Biol. Chem. 269, 18,794–18,813.Google Scholar
  11. 11.
    Shier, W. T., Lin, Y., and DeVries, A. L. (1975) Structure of the carbohydrate of antifreeze glycoproteins from an antarctic fish. FEBS Lett. 54, 135–138.CrossRefGoogle Scholar
  12. 12.
    Spiro, R. G. and Bhoyroo, V. D. (1974) Structure of O-glycosidically linked carbohydrate units of fetuin. J. Biol. Chem. 249, 5704–5717.Google Scholar
  13. 13.
    Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., and Smith, F. (1956) Colorometric method for determination of sugars and related substances. Anal. Chem. 28, 350–356.CrossRefGoogle Scholar
  14. 14.
    Ching, C. K. and Rhodes, J. M. (1990) Purification and characterisation of a pea-nut-agglutinin-binding pancreatic serum-related mucus glycoprotein. Int. J. Cancer 45, 1022–1027.CrossRefGoogle Scholar
  15. 15.
    Hounsell, E. F., Lawson, A. M., Feeney, J., Goal, H. C., Pickering, N. J., Stoll, M. S., Lui, S. C., and Feizi, T. (1985) Structural analysis of the O-glycosidically linked core-region oligosaccharides of human meconium glycoproteins which express oncofetal antigens. Eur. J. Biochem. 148, 367–377.CrossRefGoogle Scholar
  16. 16.
    Campbell, B. J., Davies, M., Rhodes, J. M., and Hounsell, E. F. (1993) Separation of neutral oligosaccharide alditols from human meconium using high pH anion exchange chromatography. J. Chromatogr. (Biomed. Appl.) 622, 137–146.CrossRefGoogle Scholar
  17. 17.
    Klein, A., Carnoy, C., Wieruszeski, J. M., Strecker, G., Strang, A. M., van Halbeek, H., Roussel, P., and Lamblin, G. (1992) The broad diversity of neutral and sialylated oligosaccharides derived from human salivary mucins. Biochemistry 31, 6152–6165.CrossRefGoogle Scholar
  18. 18.
    Chai, W., Hounsell, E. F., Cashmore, G. C., Rosankiewiez, J. R., Bauer, C. J., Feeney, J., Frizi, T., and Lawson, A. M. (1992) Neutral oligosaccharides of bovine submaxillary mucin a combined mass spectrometry and 1H-NMR study. Eur. J. Biochem. 203, 257–268.CrossRefGoogle Scholar
  19. 19.
    O’Lloyd, K. and Savage, A. (1991) High performance anion-exchange chromatography of reduced oligosaccharides from sialomucins. Glycoconj. J. 8, 493–498.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1998

Authors and Affiliations

  • Barry J. Campbell
    • 1
  • Jonathan M. Rhodes
    • 1
  1. 1.Glycobiology Group, Gastroenterology Research Unit, Department of MedicineThe University LiverpoolUK

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