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

Glycoconjugate Journal volume 8, pages 434–441 (1991)Cite this article

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.

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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)

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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
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  2. Shida Yousefi
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  3. James W. Dennis
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  4. Roland Schauer
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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

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

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