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Is Permethylation Strategy Always Applicable to Protein N-Glycosylation Study?: A Case Study on the O-Acetylation of Sialic Acid in Fish Serum Glycans

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Functional Glycomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 600))

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Abstract

O-Acetylation is one of the major modifications of sialic acids that significantly alters biological properties of the parent molecule. These O-acetylated forms are components of the cellular membrane and can affect physiological and pathological responses. Understanding the role of N-glycans in physiology is of increasing relevance to cellular biologists in various disciplines who study glycoproteomics yet lack information regarding the function of the attached glycans. However, permethylation, the most common mass spectrometric analytical means, leads to the loss of O-linked acetyl groups in sialic acids. In this chapter, we demonstrated that O-acetylation of sialic acid in Atlantic salmon serum N-glycan can be well investigated by capillary electrophoresis–mass spectrometry.

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References

  1. Turnbull, J. E. and Field, R. A., (2007) Emerging glycomics technologies. Nat. Chem. Biol. 3, 74–77.

    Article  CAS  PubMed  Google Scholar 

  2. Klein, A. and Roussel, P., (1998) O-acetylation of sialic acids. Biochimie 80, 49–57.

    Article  CAS  PubMed  Google Scholar 

  3. Schauer, R. (2000) Achievements and challenges of sialic acid research. Glycoconj. J. 17, 485–499.

    Article  CAS  PubMed  Google Scholar 

  4. Varki, A. (1992) Diversity in the sialic acids. Glycobiology 3, 25–40.

    Article  Google Scholar 

  5. Campbell, F., Appleton, M. A., Fuller, C. E., Greeff, M. P., Hallgrimsson, J., Katoh, R., Ng, O. L., Satir, A., Williams, G. T., and Williams, E. D. (1994) Racial variation in the O-acetylation phenotype of human colonic mucosa. J. Pathol. 174, 169–174.

    Article  CAS  PubMed  Google Scholar 

  6. Corfield, A. P., Wagner, S. A., Paraskeva, C., Clamp, J. R., Durdey, P., Reuter, G., and Schauer, R., (1992) Loss of sialic acid O-acetylation in human colorectal cancer cells. Biochem. Soc. Trans. 20, 94S.

    CAS  PubMed  Google Scholar 

  7. Corfield, A. P., Myerscough, N., Warren, B. F., Durdey, P., Paraskeva, C., and Schauer, R. (1999) Reduction of sialic acid O-acetylation in human colonic mucins in the adenoma-carcinoma sequence. Glycoconj. J. 16, 307–317.

    Article  CAS  PubMed  Google Scholar 

  8. Ghosh, S., Bandyopadhyay, S., Mallick, A., Pal, S., Vlasak, R., Bhattacharya, D. K., and Mandal, C. (2005) Interferon gamma promotes survival of lymphoblasts overexpressing 9-O-acetylated sialoglycoconjugates in childhood acute lymphoblastic leukaemia (ALL). J. Cell Biochem. 95, 206–216.

    Article  CAS  PubMed  Google Scholar 

  9. Ghosh, S., Bandyopadhyay, S., Mukherjee, K., Mallick, A., Pal, S., Mandal, C., Bhattacharya, D. K., and Mandal, C. (2007) O-acetylation of sialic acids is required for the survival of lymphoblasts in childhood acute lymphoblastic leukemia (ALL). Glycoconj. J. 24, 17–24.

    Article  CAS  PubMed  Google Scholar 

  10. Shen, Y., Tiralongo, J., Iwersen, M., Sipos, B., Kalthoff, H., and Schauer, R. (2002) Characterization of the sialate-7(9)-O-acetyltransferase from the microsomes of human colonic mucosa. Biol. Chem. 383, 307–317.

    Article  CAS  PubMed  Google Scholar 

  11. Shen, Y., Tiralongo, J., Kohla, G., and Schauer, R. (2004) Regulation of sialic acid O-acetylation in human colon mucosa. Biol. Chem. 385, 145–152.

    Article  CAS  PubMed  Google Scholar 

  12. Shen, Y., Kohla, G., Lrhorfi, A. L., Sipos, B., Kalthoff, H., Gerwig, G. J., Kamerling, J. P., Schauer, R., and Tiralongo, J., (2004) O-acetylation and de-O-acetylation of sialic acids in human colorectal carcinoma. Eur. J. Biochem. 271, 281–290.

    Article  CAS  PubMed  Google Scholar 

  13. Shi, W. X., Chammas, R., and Varki, A. (1996) Regulation of sialic acid 9-O-acetylation during the growth and differentiation of murine erythroleukemia cells. J. Biol. Chem. 271, 31517–31525.

    Article  CAS  PubMed  Google Scholar 

  14. Sjoberg, E. R., Powell, L. D., Klein, A., and Varki, A. (1994) Natural ligands of the B cell adhesion molecule CD22 beta can be masked by 9-O-acetylation of sialic acids. J. Cell Biol. 126, 549–562.

    Article  CAS  PubMed  Google Scholar 

  15. Butor, C., Diaz, S., and Varki, A. (1993) High level O-acetylation of sialic acids on N-linked oligosaccharides of rat liver membranes. Differential subcellular distribution of 7- and 9-O-acetyl groups and of enzymes involved in their regulation. J. Biol. Chem. 268, 10197–10206.

    CAS  PubMed  Google Scholar 

  16. Morimoto, N., Nakano, M., Kinoshita, M., Kawabata, A., Morita, M., Oda, Y., Kuroda, R., and Kakehi, K., (2001) Specific distribution of sialic acids in animal tissues as examined by LC-ESI-MS after derivatization with 1,2-diamino-4,5-methylenedioxybenzene. Anal. Chem. 73, 5422–5428.

    Article  CAS  PubMed  Google Scholar 

  17. Stehling, P., Gohlke, M., Fitzner, R., and Reutter, W. (1998) Rapid analysis of O-acetylated neuraminic acids by matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Glycoconj. J. 15, 339–344.

    Article  CAS  PubMed  Google Scholar 

  18. Hara, S., Yamaguchi, M., Takemori, Y., Furuhata, K., Ogura, H., and Nakamura, M. (1989) Determination of mono-O-acetylated N-acetylneuraminic acids in human and rat sera by fluorometric high-performance liquid chromatography. Anal. Biochem. 179, 162–166.

    Article  CAS  PubMed  Google Scholar 

  19. Higa, H. H., Manzi, A. and Varki, A., (1989) O-acetylation and de-O-acetylation of sialic acids. Purification, characterization, and properties of a glycosylated rat liver esterase specific for 9-O-acetylated sialic acids. J. Biol. Chem. 264, 19435–19442.

    CAS  PubMed  Google Scholar 

  20. Harvey, D. J., (2006) Analysis of carbohydrates and glycoconjugates by matrix-assisted laser desorption/ionization mass spectrometry: An update covering the period 1999-2000. Mass Spectrom. Rev. 25, 595–662.

    Article  CAS  PubMed  Google Scholar 

  21. Liu, X., Li, X., Chan, K., Zou, W., Pribil, P., Li, X. F., Sawyer, M. B., and Li, J. (2007) “One-pot" methylation in glycomics application: esterification of sialic acids and permanent charge construction. Anal. Chem. 79, 3894–3900.

    Article  CAS  PubMed  Google Scholar 

  22. Sekiya, S., Wada, Y. and Tanaka, K., (2005) Derivatization for stabilizing sialic acids in MALDI-MS. Anal. Chem. 77, 4962–4968.

    Article  CAS  PubMed  Google Scholar 

  23. Sagi, D., Kienz, P., Denecke, J., Marquardt, T., and Peter-Katalinic, J. (2005) Glycoproteomics of N-glycosylation by in-gel deglycosylation and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry mapping: application to congenital disorders of glycosylation. Proteomics. 5, 2689–2701.

    Article  CAS  PubMed  Google Scholar 

  24. Ylonen, A., Kalkkinen, N., Saarinen, J., Bogwald, J., and Helin, J. (2001) Glycosylation analysis of two cysteine proteinase inhibitors from Atlantic salmon skin: di-O-acetylated sialic acids are the major sialic acid species on N-glycans. Glycobiology 11, 523–531.

    Article  CAS  PubMed  Google Scholar 

  25. Ylonen, A., Helin, J., Bogwald, J., Jaakola, A., Rinne, A., and Kalkkinen, N., (2002) Purification and characterization of novel kininogens from spotted wolffish and Atlantic cod. Eur. J. Biochem. 269, 2639–2646.

    Article  CAS  PubMed  Google Scholar 

  26. Liu, X., Afonso, L., Altman, E., Johnson, S., Brown, L., and Li, J. (2008) O-acetylation of sialic acids in N-glycans of Atlantic salmon (Salmo salar) serum is altered by handling stress. Proteomics 8, 2849–2857.

    Article  CAS  PubMed  Google Scholar 

  27. Kang, P., Mechref, Y., Kyselova, Z., Goetz, J. A., and Novotny, M. V. (2007) Comparative glycomic mapping through quantitative permethylation and stable-isotope labeling. Anal. Chem. 79, 6064–6073.

    Article  CAS  PubMed  Google Scholar 

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Liu, X., Afonso, L. (2010). Is Permethylation Strategy Always Applicable to Protein N-Glycosylation Study?: A Case Study on the O-Acetylation of Sialic Acid in Fish Serum Glycans. In: Li, J. (eds) Functional Glycomics. Methods in Molecular Biology, vol 600. Humana Press. https://doi.org/10.1007/978-1-60761-454-8_18

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  • DOI: https://doi.org/10.1007/978-1-60761-454-8_18

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-453-1

  • Online ISBN: 978-1-60761-454-8

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