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Composition of N-linked carbohydrates from ovalbumin and co-purified glycoproteins

  • D. J. Harvey
  • D. R. Wing
  • B. Küster
  • I. B. H. Wilson
Article

Abstract

Analysis of commercial samples of chicken ovalbumin by reversed-phase high performance liquid chromatography and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) showed the presence of several other co-purifying glycoproteins. These were isolated, subjected to tryptic digestion, and two of them were identified as ovomucoid and chicken riboflavin binding-protein following database matching of the peptide masses obtained by MALDI. The N-linked glycans were released from the glycoproteins and their structures were examined by MALDI-MS in combination with exoglycosidase digestion. Ovalbumin was found to be glycosylated mainly with high-mannose and hybrid structures, consistent with profiles obtained on the intact glycoprotein by electrospray. The other glycoproteins contained mainly larger, complex glycans with up to five antennae, many of which had earlier been associated with ovalbumin.

Keywords

High Performance Liquid Chromatography GlcNAc Sinapinic Acid Glycan Profile Complex Glycan 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Neuberger, A. Carbohydrates in proteins I. The carbohydrate component of crystalline egg albumin. Biochem. J. 1938, 32, 1435–1451.Google Scholar
  2. 2.
    Huang, C.-C.; Mayer, H. E., Jr.; Montgogery, R. Microheterogeneity and paucidispersity of glycoproteins, Part 1. The carbohydrate of chicken ovalbumin. Carbohydrate Res. 1970, 13, 127–137.CrossRefGoogle Scholar
  3. 3.
    Tai, T.; Yamishita, K.; Ogata-Arakawa, M.; Koide, N.; Muramatsu, T.; Iwashita, S.; Inoue, Y.; Kobata, A. Structural studies of two ovalbumin glycopeptides in relation to the endo-β-N-acetyl-glucosaminidase specificity. J. Biol. Chem. 1975, 250, 8569–8575.Google Scholar
  4. 4.
    Tai, T.; Yamishita, K.; Ito, S.; Kobata, A. Structures of the carbohydrate moiety of ovalbumin glycopeptide III and the difference in specificity of endo-β-N-acetylglucosaminidases C11 and H. J. Biol. Chem. 1977, 252, 6687–6694.Google Scholar
  5. 5.
    Yamashita, K.; Tachibana, Y.; Kobata, A. The structures of the galactose-containing sugar chains of ovalbumin. J. Biol. Chem. 1978, 253, 3862–3869.Google Scholar
  6. 6.
    Yamashita, K.; Ueda, I.; Kobata, A. Sulfated asparagine-linked sugar chains of hen egg albumin J. Biol. Chem. 1983, 258, 14144–14147.Google Scholar
  7. 7.
    Yamashita, K.; Tachibana, Y.; Hitoi, A.; Kobata, A. Sialic acid-containing sugar chains of hen ovalbumin and ovomucoid. Carbohydrate Res. 1984, 130, 271–288.CrossRefGoogle Scholar
  8. 8.
    Ceccarini, C.; Lorenzoni, P.; Atkinson, P. H. Fractionation of ovalbumin glycopeptide AC-C by high pressure liquid chromatography. Determination of structure by 1H-NMR spectroscopy. Biochim. Biophys. Acta 1983, 759, 214–221.Google Scholar
  9. 9.
    Nomoto, H.; Inoue, Y. A novel glycoasparagine isolated from an ovalbumin glycopeptide fraction (GP-IV). Anion-exchange borate chromatography and structural analysis of GP-IV glycoasparagines. Eur. J. Biochem. 1983, 135, 243–250.CrossRefGoogle Scholar
  10. 10.
    Rago, R. P.; Ramirez-Soto, D.; Poretz, R. D. Two-dimensional poly(acrylamide) electrophoresis of fluresceinated glycopeptides. Resolution and structural characterization of ovalbumin glycans. Carbohydrate Res. 1992, 236, 1–8.CrossRefGoogle Scholar
  11. 11.
    Duffin, K. L.; Welply, J. K.; Huang, E.; Henion, J. D. Characterization of N-linked oligosaccharides by electrospray and tandem mass spectrometry. Anal. Chem. 1992, 64, 1440–1448.CrossRefGoogle Scholar
  12. 12.
    Harvey, D. J.; Rudd, P. M.; Bateman, R. H.; Bordoli, R. S.; Howes, K.; Hoyes, J. B.; Vickers, R. G. Examination of complex oligosaccharides by matrix-assisted laser desorption/ionization mass spectrometry on time-of-flight and magnetic sector instruments. Org. Mass Spectrom. 1994, 29, 753–765.CrossRefGoogle Scholar
  13. 13.
    Bordoli, R. S.; Howes, K.; Vickers, R. G.; Bateman, R. H.; Harvey, D. J. Matrix-assisted laser desorption mass spectrometry on a magnetic sector instrument fitted with an array detector. Rapid Commun. Mass Spectrom. 1994, 8, 585–589.CrossRefGoogle Scholar
  14. 14.
    Bordoli, R. S.; Howes, K.; Vickers, R. G.; Bateman, R. H.; Harvey, D. J. Matrix-assisted laser desorption mass spectrometry on a magnetic sector instrument fitted with an array detector. Mass Spectrom. Biomol. Sci., NATO ASI Ser. C 1996, 475, 51–60.Google Scholar
  15. 15.
    Da Silva, M. L. C.; Stubbs, H. J.; Tamura, T.; Rice, K. G. 1H NMR characterization of a hen ovalbumin tyrosinamide N-linked oligosaccharide library. Arch. Biochem. Biophys. 1995, 318, 465–475.CrossRefGoogle Scholar
  16. 16.
    Küster, B.; Wheeler, S. F.; Hunter, A. P.; Dwek, R. A.; Harvey, D. J. Sequencing of N-linked oligosaccharides directly from protein gels: In-gel deglycosylation followed by matrix-assisted laser desorption/ionization mass spectrometry and normal-phase high-performance liquid chromatography. Anal. Biochem. 1997, 250, 82–101.CrossRefGoogle Scholar
  17. 17.
    North, S.; Okafo, G.; Birrell, H.; Haskins, N.; Camilleri, P. Minimizing cationization effects in the analysis of complex mixtures of oligosaccharides. Rapid Commun. Mass Spectrom. 1997, 11, 1635–1642.CrossRefGoogle Scholar
  18. 18.
    North, S.; Birrell, H.; Camilleri, P. Positive and negative ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of complex glycans released from hen ovalbumin and derivatized with 2-aminoacridone. Rapid Commun. Mass Spectrom. 1998, 12, 349–356.CrossRefGoogle Scholar
  19. 19.
    Anumula, K. R.; Dhume, S. T. High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivatization with highly fluorescent anthranilic acid. Glycobiology 1998, 8, 685–694.CrossRefGoogle Scholar
  20. 20.
    Mechref, Y.; Novotny, M. V. Mass spectrometric mapping and sequencing of N-linked oligosaccharides derived from submicrogram amounts of glycoproteins. Anal. Chem. 1998, 70, 455–463.CrossRefGoogle Scholar
  21. 21.
    Stevens, L. Egg white proteins. Comp. Biochem. Physiol. 1991, 100B, 1–9.Google Scholar
  22. 22.
    Patel, T.; Bruce, J.; Merry, A.; Bigge, C.; Wormald, M.; Jaques, A.; Parekh, R. Use of hydrazine to release in intact and unreduced form both N- and O-linked oligosaccharides from glycoproteins. Biochemistry 1993, 32, 679–693.CrossRefGoogle Scholar
  23. 23.
    Harvey, D. J. Quantitative aspects of the matrix-assisted laser desorption mass spectrometry of complex oligosaccharides. Rapid Commun. Mass Spectrom. 1993, 7, 614–619.CrossRefGoogle Scholar
  24. 24.
    Piskarev, V. E.; Sepetov, N. F.; Likhosherstov, L. M.; Galenko, E. L.; Derevitskaia, V. A. Structure of carbohydrate chains of the riboflavin-binding glycoprotein of chicken egg protein. II. 1H-NMR (500 MHz) spectroscopy of the major neutral oligosaccharides. Bioorg. Khim. 1989, 15, 156–1554.Google Scholar
  25. 25.
    Yamashita, K.; Kamerling, J. P.; Kobata, A. Structural studies of the sugar chains of hen ovomucoid: Evidence indicating that they are formed mainly by the alternate biosynthetic pathway of asparagine-linked sugar chains. J. Biol. Chem. 1983, 258, 3099–3106.Google Scholar
  26. 26.
    Yamashita, K.; Kamerling, J. P.; Kobata, A. Structural study of the carbohydrate moiety of hen ovomucoid: Occurrence of a series of pentaantennary complex-type asparagine-linked sugar chains. J. Biol. Chem. 1982, 257, 12809–12814.Google Scholar
  27. 27.
    Paz-Parente, J.; Strecker, G.; Leroy, Y.; Montreuil, J.; Fournet, B.; van-Halbeek, H.; Dorland, L.; Vliegenthart, J. F. Primary structure of a novel N-glycosidic carbohydrate unit, derived from hen ovomucoid. A 500-MHz 1H-NMR study. FEBS Lett. 1983, 152, 145–152.CrossRefGoogle Scholar
  28. 28.
    Egge, H.; Peter-Katalinic, J.; Paz-Parente, J.; Strecker, G.; Montreuil, J.; Fournet, B. Carbohydrate structures of hen ovomucoid. A mass spectrometric analysis. FEBS Lett. 1983, 156, 357–362.CrossRefGoogle Scholar

Copyright information

© American Society for Mass Spectrometry 2000

Authors and Affiliations

  • D. J. Harvey
    • 1
  • D. R. Wing
    • 1
  • B. Küster
    • 1
  • I. B. H. Wilson
    • 1
  1. 1.Department of BiochemistryOxford Glycobiology InstituteOxfordUK

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