Skip to main content
SpringerLink
Log in

Two closely related forms of UDP-GlcNAc: α6-D-mannoside β1,2-N-acetylglucosaminyltransferase II occur in the clawed frog Xenopus laevis

  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

UDP-GlcNAc:α6-D-mannoside β1,2-N-acetylglucosaminyltransferase II (GnT II; EC 2.4.1.143) is a medial-Golgi resident enzyme that catalyses an essential step in the biosynthetic pathway leading from high mannose to complex N-linked oligosaccharides. Screening a cDNA library from Xenopus laevis ovary with a human GnT II DNA probe resulted in the isolation of two cDNA clones encoding two closely related GnT II isoenzymes, GnT II-A and GnT II-B. Analysis of the corresponding genomic DNAs revealed that the open reading frame of both X. laevis GnT II genes resides within a single exon. The GnT II-A gene was found to be transcriptionally active in all X. laevis tissues tested. In contrast, expression of the GnT II-B gene was detected only in a limited number of tissues. Both GnT II-A and GnT II-B exhibit a type II transmembrane protein topology with a putative N-terminal cytoplasmic tail of 9 amino acids followed by a transmembrane domain of 18 residues, and a C-terminal luminal domain of 405 residues. The two proteins differ at 28 amino acid positions within their luminal regions. Heterologous expression of soluble forms of the enzymes in insect cells showed that GnT II-A and GnT II-B are both catalytically active and exhibit similar specific activities. Both recombinant proteins are modified with N-linked oligosaccharides. N-terminal deletion studies demonstrated that the first 49 amino acid residues are not essential for proper folding and enzymatic activity of X. laevis GnT II. Published in 2003.

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

Similar content being viewed by others

References

  1. Schachter H, Biochem Cell Biol 64, 163–81 (1986).

    Google Scholar 

  2. Kornfeld R, Kornfeld S, Annu Rev Biochem 54, 631–44 (1985).

    Google Scholar 

  3. Charuk JHM, Tan J, Bernardini M, Haddad S, Reithmeier RAF, Jaeken J, Schachter H, Eur J Biochem 230, 797–805 (1995).

    Google Scholar 

  4. Jaeken J, Schachter H, Carchon H, De Cock P, Coddeville B, Spik G, Arch Dis Child 71, 123–7 (1994).

    Google Scholar 

  5. Jaeken J, Spik G, Schachter H, Glycoproteins and Disease (Elsevier, Amsterdam, The Netherlands, 1996), pp. 457–67.

    Google Scholar 

  6. Tan J, Dunn J, Jaeken J, Schachter H, Am J Hum Genet 59, 810–7 (1996).

    Google Scholar 

  7. Schachter H, Jaeken J, Biochim Biophys Acta 1455, 179–92 (1999).

    Google Scholar 

  8. Wang Y, Tan J, Campbell RM, Ditto D, Le D, Schachter H, Marth JD, Glycobiology 10, 1131–2 (2000).

    Google Scholar 

  9. Wang Y, Tan J, Sutton-Smith M, Ditto D, Panico M, Campbell RM, Varki NM, Long JM, Jaeken J, Levison SR, Wynshaw BA, Morris HR, Le D, Dell A, Schachter H, Mart JD, Glycobiology 11, 1051–70 (2001).

    Google Scholar 

  10. Tan J, D'Agostaro GAF, Bendiak B, Reck F, Sarkar M, Squire JA, Leong P, Schachter H, Eur J Biochem 231, 317–28 (1995).

    Google Scholar 

  11. D'Agostaro GAF, Zingoni A, Moritz RL, Simpson RJ, Schachter H, Bendiak BJ, J Biol Chem 270, 15211–21 (1995).

    Google Scholar 

  12. Campbell R, Tan J, Schachter H, Bendiak B, Marth J, Glycobiology 7, 1050 (1997).

    Google Scholar 

  13. Leeb T, Kriegesmann B, Baumgartner BG, Klett C, Yerle M, Hameister H, Brenig B, Biochim Biophys Acta 1336, 361–6 (1997).

    Google Scholar 

  14. Chen S, Tan J, Reinhold VN, Spence AM, Schachter H, Biochim Biophys Acta 1573, 271–9 (2002).

    Google Scholar 

  15. Strasser R, Steinkellner H, Boren M, Altmann F, Mach L, Glössl J, Mucha J, Glycoconjugate J 16, 787–91 (1999).

    Google Scholar 

  16. Santacruz-Toloza L, Huang Y, John SA, Papazian, DM, Biochemistry 33, 5607–13 (1994).

    Google Scholar 

  17. Khanna R, Myers MP, Laine M, Papazian DM, J Biol Chem 276, 34028–34 (2001).

    Google Scholar 

  18. Roitsch T, Lehle L, Eur J Biochem 181, 733–9 (1989).

    Google Scholar 

  19. Cantor AB, Kornfeld S, Anal Biochem 205, 220–6 (1992).

    Google Scholar 

  20. Mucha J, Svoboda B, Fröhwein U, Strasser R, Mischinger M, Schwihla H, Altmann F, Hane W, Schachter H, Glössl J, Mach L, Glycobiology 11, 769–78 (2001).

    Google Scholar 

  21. Sarkar M, Pagny S, Unligil UM, Joziasse D, Mucha J, Glössl J, Schachter H, Glycoconjugate J 15, 193–7 (1998).

    Google Scholar 

  22. Chomczynski P, Sacchi N, Anal Biochem 162, 156–9 (1987).

    Google Scholar 

  23. Ausubel FM et al. (ed.), Current Protocols in Molecular Biology (John Wiley & Sons, Inc., 1994‐2002), Vol. 1, Ch. 2.

  24. Unligil UM, Zhou S, Yuwaraj S, Sarkar M, Schachter H, Rini JM, EMBO J 19, 5269–80 (2000).

    Google Scholar 

  25. Altmann F, Anal Biochem 204, 215–9 (1992).

    Google Scholar 

  26. Altmann F, Kornfeld G, Dalik T, Staudacher E, Glössl J, Glycobiology 3, 619–25 (1993).

    Google Scholar 

  27. Kyte J, Doolittle RF, J Mol Biol 157, 105–32 (1982).

    Google Scholar 

  28. Breton C, Mucha J, Jeanneau C, Biochimie 83, 713–8 (2001).

    Google Scholar 

  29. Wiggins CAR, Munro S, Proc Natl Acad Sci USA 95, 7945–50 (1998).

    Google Scholar 

  30. Bisbee CA, Baker MA, Wilson AC, Haji-Azimi I, Fischberger M, Science 195, 785–7 (1977).

    Google Scholar 

  31. Amaya E, Offield MF, Grainger RM, Trends Genet 14, 253–5 (1998).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Zentrum für Angewandte Genetik, Universität für Bodenkultur Wien, Muthgasse 18, A-1190, Wien, Austria

    Jan Mucha, Barbara Svoboda, Sonja Kappel, Richard Strasser, Peter Bencur, Ulrike Fröhwein, Lukas Mach & Josef Glössl

  2. Program in Structural Biology and Biochemistry, The Hospital for Sick Children, Toronto, M5G 1X8, Canada

    Harry Schachter

Authors
  1. Jan Mucha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Barbara Svoboda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sonja Kappel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Richard Strasser
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peter Bencur
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ulrike Fröhwein
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Harry Schachter
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lukas Mach
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Josef Glössl
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lukas Mach.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mucha, J., Svoboda, B., Kappel, S. et al. Two closely related forms of UDP-GlcNAc: α6-D-mannoside β1,2-N-acetylglucosaminyltransferase II occur in the clawed frog Xenopus laevis . Glycoconj J 19, 187–195 (2002). https://doi.org/10.1023/A:1024201824354

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1024201824354

Search

Navigation