Advertisement

Glycoconjugate Journal

, 26:935 | Cite as

Assays of sialate-O-acetyltransferases and sialate-O-acetylesterases

  • G. Vinayaga Srinivasan
  • Roland Schauer
Article

Abstract

The O-acetylation of sialic acids is one of the most frequent modifications of these monosaccharides and modulates many cell biological and pathological events. Sialic acid-specific O-acetyltransferases and O-acetylesterases are responsible for the metabolism of esterified sialic acids. Assays were developed for the analysis of the activities and specificities of these enzymes. The methods had to be varied in dependence on the substrate assayed, the kind of biological source, and the state of enzyme purity. With the new techniques the primary site of O-acetyl incorporation at C-7, catalyzed by the animal sialate-O-acetyltransferases studied, was ascertained. Correspondingly, this enzyme, for example from bovine submandibular gland, can be denominated as AcCoA:sialate-7-O-acetyltransferase (EC 2.3.1.45). Methods for assaying the activity of esterases de-O-acetylating sialic acids and their metabolic cooperation with the O-acetyltransferases are presented.

Keywords

Sialic acids O-acetylated O-Acetyltransferases O-Acetylesterases Thin-layer chromatography High-performance liquid chromatography Radiometry Colorimetry 

Notes

Acknowledgments

We thank Dr. Reinhard Schwartz-Albiez and Dirk Wipfler, Heidelberg, for a sample of human lymphocytes, Dr. Reinhard Vlasak, Salzburg, for influenza C viruses, Dr. Bernhard Kniep, Dresden, for O-acetylated GD3 and Dr. Guido Kohla, Hannover, for technical advice. This work was performed with a grant from Deutsche Forschungsgemeinschaft (Scha 202/33-1) and support by the Sialic Acids Society, Kiel.

References

  1. 1.
    Angata, T., Varki, A.: Chemical diversity in the sialic acids and related a-keto acids: an evolutionary perspective. Chem. Rev. 102, 439–469 (2002)CrossRefPubMedGoogle Scholar
  2. 2.
    Tiralongo, J., Schauer, R.: The enigma of enzymatic sialic acid O-acetylation. Trends Glycosci. Glycotechnol. 16, 1–15 (2004)Google Scholar
  3. 3.
    Schauer, R.: Sialic acids: Fascinating sugars in higher animals and man. Zoology 107, 49–64 (2004)CrossRefPubMedGoogle Scholar
  4. 4.
    Schauer, R., Kamerling, J.P.: Chemistry, biochemistry and biology of sialic acids. In: Montreuil, J., Vliegenthart, J.F.G., Schachter, H. (eds.) Glycoproteins II, pp. 243–402. Elsevier, Amsterdam (1997)CrossRefGoogle Scholar
  5. 5.
    Iwersen, M., Vandamme-Feldhaus, V., Schauer, R.: Enzymatic 4-O-acetylation of N-acetylneuraminic acid in guinea-pig liver. Glycoconj. J. 15, 895–904 (1998)CrossRefPubMedGoogle Scholar
  6. 6.
    Iwersen, M., Dora, H., Kohla, G., Gasa, S., Schauer, R.: Solubilisation and properties of the sialate-4-O-acetyltransferase from guinea pig liver. Biol. Chem. 384, 1035–1047 (2003)CrossRefPubMedGoogle Scholar
  7. 7.
    Higa, H.H., Butor, C., Diaz, S., Varki, A.: O-Acetylation and de-O-acetylation of sialic acids. O-Acetylation of sialic acids in rat liver Golgi apparatus involves an acetyl intermediate and essential histidine and lysine residues: a trans-membrane reaction. J. Biol. Chem. 264, 19427–19434 (1989)PubMedGoogle Scholar
  8. 8.
    Shen, Y., Kohla, G., Lrhorfi, A.L., Sipos, B., Kalthoff, H., Gerwig, G.J., Kamerling, J.P., Schauer, R., Tiralongo, J.: O-Acetylation and de-O-acetylation of sialic acids in human colorectal carcinoma. Eur. J. Biochem. 271, 281–290 (2004)CrossRefPubMedGoogle Scholar
  9. 9.
    Lrhorfi, L.A., Srinivasan, G.V., Schauer, R.: Properties and partial purification of sialate-O-acetyltransferase from bovine submandibular glands. Biol. Chem. 388, 297–306 (2007)CrossRefPubMedGoogle Scholar
  10. 10.
    Houliston, R.C., Endtz, H.P., Yuki, N., Li, J., Jarrell, H.C., Koga, M., van Belkum, A., Karwaski, M.-F., Wakarchuk, W.W., Gilbert, M.: Identification of a sialate O-acetyltransferase from Campylobacter jejuni. J. Biol. Chem. 281, 11480–11486 (2006)CrossRefPubMedGoogle Scholar
  11. 11.
    Claus, H., Borrow, R., Achtman, M., Morelli, G., Kantelberg, C., Longworth, E., Frosch, M., Vogel, U.: Genetics of capsule O-acetylation in serogroup C, W-135 and Y meningococci. Mol. Microbiol. 51, 227–239 (2004)CrossRefPubMedGoogle Scholar
  12. 12.
    Steenbergen, S.M., Lee, Y.-C., Vann, W.F., Vionnet, J., Wright, L.F., Vimr, E.R.: Separate pathways for O-Acetylation of polymeric and monomeric sialic acids and identification if sialyl O-acetylesterase in Escherichia coli K1. J. Bacteriol. 188, 6195–6206 (2006)CrossRefPubMedGoogle Scholar
  13. 13.
    Lewis, A.L., Nizet, V., Varki, A.: Discovery and characterization of sialic acid O-acetylation in group B Streptococcus. Proc. Natl. Acad. Sci. USA 101, 11123–11128 (2004)CrossRefPubMedGoogle Scholar
  14. 14.
    Fahr, C., Schauer, R.: Detection of sialic acids and gangliosides with special reference to 9-O-acetylated species in basaliomas and normal human skin. J. Invest. Dermatol. 116, 254–260 (2001)CrossRefPubMedGoogle Scholar
  15. 15.
    Hubl, U., Ishida, H., Kiso, M., Hasegawa, A., Schauer, R.: Studies on the specificity and sensitivity of the influenza C virus binding assay for O-acetylated sialic acids and its application to human melanomas. J. Biochem. 127, 1021–1031 (2000)PubMedGoogle Scholar
  16. 16.
    Sinha, D., Mandal, C., Bhattacharya, D.K.: Identification of 9-O acetyl sialoglycoconjugates (9-OAcSGs) as biomarkers in childhood acute lymphoblastic leukemia using a lectin, AchatininH, as a probe. Leukemia 13, 119–125 (1999)CrossRefPubMedGoogle Scholar
  17. 17.
    Shen, Y., Tiralongo, J., Kohla, G., Schauer, R.: Regulation of sialic acid O-acetylation in human colon mucosa. Biol. Chem. 385, 145–152 (2004)CrossRefPubMedGoogle Scholar
  18. 18.
    Higa, H.H., Manzi, A., Varki, A.: O-Acetylation and de-O-acetylation of sialic acids. J. Biol. Chem. 264, 19435–19442 (1989)PubMedGoogle Scholar
  19. 19.
    Lewis, A.L., Cao, H., Patel, S.K., Diaz, S., Ryan, W., Carlin, A.F., Thon, V., Lewis, W.G., Varki, A., Chen, X., Nizet, V.: NeuA sialic acid O-acetylesterase activity modulates O-acetylation of capsular polysaccharide in group B Streptococcus. J. Biol. Chem. 282, 27562–27571 (2007)CrossRefPubMedGoogle Scholar
  20. 20.
    Schauer, R., Reuter, G., Stoll, S., Shukla, A.K.: Partial purification and characterization of sialate O-acetylesterase from bovine brain. J. Biochem. 106, 143–150 (1989)PubMedGoogle Scholar
  21. 21.
    Higa, H.H., Manzi, A., Diaz, S., Varki, A.: Sialate-9-O-acetylesterase from rat liver. Methods Enzymol. 179, 409–415 (1989)CrossRefPubMedGoogle Scholar
  22. 22.
    Strasser, P., Unger, U., Strobl, B., Vilas, U., Vlasak, R.: Recombinant viral sialate-O-acetylesterases. Glycoconj. J. 20, 551–561 (2004)CrossRefPubMedGoogle Scholar
  23. 23.
    Schauer, R., Shukla, A.K.: Isolation and properties of two sialate-O-acetylesterases from horse liver with 4- and 9-O-acetyl specificities. Glycoconj. J. (2008; in press). doi:  10.1007/s10719-008-9109-9
  24. 24.
    Smits, S.L., Gerwig, G.J., van Vliet, A.L.W., Lissenberg, A., Briza, P., Kamerling, J.P., Vlasak, R., de Groot, R.J.: Nidovirus sialate-O-acetylesterases. J. Biol. Chem. 280, 6933–6941 (2005)CrossRefPubMedGoogle Scholar
  25. 25.
    Vandamme, V., Cazlaris, H., Le Marer, N., Laudet, V., Lagrou, C., Verbert, A., Delannoy, P.: Comparison of sialyl and a-1,3-galactosyltransferase activity in NIH3T3 cells transformed with ras oncogene: increased b-galactoside a-2,6-sialyltransferase. Biochimie 74, 89–100 (1992)CrossRefPubMedGoogle Scholar
  26. 26.
    Diaz, S., Higa, H.H., Hayes, B.K., Varki, A.: O-Acetylation and de-O-acetylation of sialic acids. J. Biol. Chem. 264, 19416–19426 (1989)PubMedGoogle Scholar
  27. 27.
    Reuter, G., Schauer, R.: Determination of sialic acids. Methods Enzymol. 230, 168–199 (1994)CrossRefPubMedGoogle Scholar
  28. 28.
    Mawhinney, T.P., Chance, D.L.: Hydrolysis of sialic acids and O-acetylated sialic acids with propionic acid. Anal. Biochem. 223, 164–167 (1994)CrossRefPubMedGoogle Scholar
  29. 29.
    Vandamme-Feldhaus, V., Schauer, R.: Characterization of the enzymatic 7-O-acetylation of sialic acids and evidence for enzymatic O-acetyl migration from C-7 to C-9 in bovine submandibular gland. J. Biochem. 124, 111–121 (1998)PubMedGoogle Scholar
  30. 30.
    Varki, A., Diaz, S.: The release and purification of sialic acids from glycoconjugates: methods to minimize the loss and migration of O-acetyl groups. Anal. Biochem. 137, 236–247 (1984)CrossRefPubMedGoogle Scholar
  31. 31.
    Hara, S., Takemori, Y., Yamaguchi, M., Nakamura, M., Ohkura, Y.: Fluorometric high-performance liquid chromatography of N-acetyl- and N-glycolylneuraminic acids and its application to their microdetermination in human and animal sera, glycoproteins, and glycolipids. Anal. Biochem. 164, 138–145 (1987)CrossRefPubMedGoogle Scholar
  32. 32.
    Dzieciatkowska, M., Brochu, D., van Belkum, A., Heikema, A.P., Yuki, N., Houliston, R.S., Richards, J.C., Gilbert, M., Li, J.: Mass spectrometric analysis of intact lipooligosaccharide: Direct evidence for O-acetylated sialic acids and discovery of O-linked glycine expressed by Campylobacter jejuni. Biochemistry 46, 14704–14714 (2007)CrossRefPubMedGoogle Scholar
  33. 33.
    Bergfeld, A.K., Claus, H., Vogel, U., Mühlenhoff, M.: Biochemical characterization of the polysialic acid specific O-acetyltransferase NeuO of Escherichia coli. J. Biol. Chem. 282, 22217–22227 (2007)CrossRefPubMedGoogle Scholar
  34. 34.
    Kamerling, J.P., Schauer, R., Shukla, A.K., Stoll, S., van Halbeek, H., Vliegenthart, J.F.G.: Migration of O-acetyl groups in N,O-acetylneuraminic acids. Eur. J. Biochem. 162, 601–607 (1987)CrossRefPubMedGoogle Scholar
  35. 35.
    Reuter, G., Pfeil, R., Stoll, S., Schauer, R., Kamerling, J.P., Versluis, C., Vliegenthart, J.F.G.: Identification of new sialic acids derived from glycoproteins of bovine submandibular gland. Eur. J. Biochem. 134, 139–143 (1983)CrossRefPubMedGoogle Scholar
  36. 36.
    Shi, W.-X., Chammas, R., Varki, A.: Linkage-specific action of endogenous sialic acid O-acetyltransferase in Chinese hamster ovary cells. J. Biol. Chem. 271, 15130–15138 (1996)CrossRefPubMedGoogle Scholar
  37. 37.
    Chen, H.Y., Challa, A.K., Varki, A.: 9-O-Acetylation of exogenously added ganglioside GD3. J. Biol. Chem. 281, 7825–7833 (2006)CrossRefPubMedGoogle Scholar
  38. 38.
    Schauer, R., Shukla, A., Srinivasan, G.V.: Functional modulation of sialoglycoconjugates by sialic acid O-acetyltransferases and esterases. Glycoconj. J. 24, 293–294 (2007)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  1. 1.Biochemisches InstitutChristian-Albrechts-UniversitätKielGermany

Personalised recommendations