Skip to main content
SpringerLink
Log in

CMP-N-acetylneuraminic acid hydroxylase activity determines the wheat germ agglutinin-binding phenotype in two mutants of the lymphoma cell line MDAY-D2

  • Papers
  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

The dominant glycosylation mutants of MDAY-D2 mouse lymphoma cells, designated class 2 (D33W25 and D34W25) were selected for their resistance to the toxic effects of wheat germ agglutinin (WGA) and shown to express elevated levels of Neu5Gc. In accordance with this, the activity of CMP-Neu5Ac hydroxylase was found to be substantially higher in the mutant cells. The hydroxylase in the D33W25 mutant cells exhibited kinetic properties identical to those of the same enzyme from mouse liver. Growth rate experimentsin vivo andin vitro, where the mutant cells grew more slowly at low cell densities in serum-free medium and also formed slower growing tumours in syngeneic mice, indicate that CMP-Neu5Ac hydroxylase expression may be associated with altered growth of the mutant cells.

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

Abbreviations

WGA:

wheat germ agglutinin

Neu5Ac:

N-acetyl-β-d-neuraminic acid

Neu5Gc:

N-glycology-β-d-neuraminic acid

CMP-Neu5Ac:

cytidine-5′-monophospho-N-acetylneuraminic acid

CMP-Neu5Gc:

cytidine-5′-monophospho-N-glycoloylneuraminic acid

FACS:

fluorescence-activated cell sorting

buffer A:

triethylamine hydrogen carbonate, pH 7.6 (concentration given at appropriate points in the text)

SFM:

serum free medium

IMDM:

Iscove's modified Dulbecco's medium

CMP-Neu5Ac hydroxylase:

CMP-N-acetylneuraminate: NAD(P)H oxido-reductase (N-acetyl hydroxylating) (EC 1.14.99.18); CMP-sialate hydrolase (EC 3.1.4.40); sialic acid-pyruvate lyase (EC 4.1.3.3)

References

  1. Dennis JW (1986)Cancer Res 46:4594–600.

    Google Scholar 

  2. Dennis JW, Laferté S, Waghorn C, Breitman ML, Kerbel RS (1987)Science 236:582–5.

    Google Scholar 

  3. Dennis JW, Laferté S, Yagel S, Breitman M (1989)Cancer Cells 1:87–92.

    Google Scholar 

  4. Peters BP, Ebisu S, Goldstein IJ, Flashner M (1979)Biochemistry 18:5505–11.

    Google Scholar 

  5. Shaw L, Schauer R (1988)Biol Chem Hoppe-Seyler 369:477–86.

    Google Scholar 

  6. Shaw L, Schauer R (1989)Biochem J 263:355–63.

    Google Scholar 

  7. Nöhle U, Shukla AK, Schröder C, Reuter G, Schauer R, Kamerling JP, Vliegenthart JFG (1985)Eur J Biochem 152:459–63.

    Google Scholar 

  8. Schauer R (1982)Adv Carbohydr Chem Biochem 40:131–234.

    Google Scholar 

  9. Nöhle U, Beau J-M, Schauer R (1982)Eur J Biochem 126:543–8.

    Google Scholar 

  10. Kerbel RS, Florian MS, Man S, Dennis JW, McKenzie I (1980)J Natl Cancer Inst 64:1221–30.

    Google Scholar 

  11. Lagarde AE, Donaghue TP, Kerbel RS, Siminovitch L (1984)Somatic Cell Molec Genet 10:503–19.

    Google Scholar 

  12. Beau J-M, Schauer R, Haverkamp J, Kamerling JP, Dorland L, Vliegenthart JFG (1984)Eur J Biochem 140:203–8.

    Google Scholar 

  13. Schauer R (1987)Methods Enzymol 138:132–61.

    Google Scholar 

  14. van der Elst I, Dennis JW (1991)Exp Cell Res 192:612–21.

    Google Scholar 

  15. Rodeck U, Herlyn M, Menssen HD, Furlanetto RW, Koprowski H (1987)Int J Cancer 40:687–90.

    Google Scholar 

  16. Chadwick DE, Lagarde AE (1988)J Natl Cancer Inst 80:318–25.

    Google Scholar 

  17. Dennis JW, Kosh K, Yousefi S, van der Elst I (1990)Cancer Res 50:1867–72.

    Google Scholar 

  18. Monsigny M, Roche A-C, Sene C, Maget-Dana R, Delmotte F (1980)Eur J Biochem 104:147–53.

    Google Scholar 

  19. Maget-Dana R, Veh RW, Sander M, Roche A-C, Schauer R, Monsigny M (1981)Eur J Biochem 114:11–6.

    Google Scholar 

  20. Lepers A, Shaw L, Schneckenburger P, Cacan R, Verbert A, Schauer R (1990)Eur J Biochem 193:715–23.

    Google Scholar 

  21. Bouhours JF, Bouhours D (1989)J Biol Chem 264:16992–99.

    Google Scholar 

  22. Howell KE, Devaney E, Gruenburg J (1989)Trends Biochem Sci 44:44–7.

    Google Scholar 

  23. Muchmore EA, Milewski M, Varki A, Diaz S (1989)J Biol Chem 264:20216–23.

    Google Scholar 

  24. Schauer R (1978)Methods Enzymol 50:374–86.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biochemisches Institut, Christian-Albrechts Universität, Olshausenstr, 40, 2300, Kiel 1, Germany

    Lee Shaw & Roland Schauer

  2. Samuel Lunenfeld Research Institute, 600 University Avenue, M5G 1X5, Toronto, Ontario, Canada

    Shida Yousefi & James W. Dennis

  3. Department of Medical Genetics, University of Toronto, Toronto, Ontario, Canada

    James W. Dennis

Authors
  1. Lee Shaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shida Yousefi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. James W. Dennis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Roland Schauer
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shaw, L., Yousefi, S., Dennis, J.W. et al. CMP-N-acetylneuraminic acid hydroxylase activity determines the wheat germ agglutinin-binding phenotype in two mutants of the lymphoma cell line MDAY-D2. Glycoconjugate J 8, 434–441 (1991). https://doi.org/10.1007/BF00731295

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00731295

Keywords

Search

Navigation