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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wheat germ agglutinin
fluorescence-activated cell sorting
- buffer A:
triethylamine hydrogen carbonate, pH 7.6 (concentration given at appropriate points in the text)
serum free medium
Iscove's modified Dulbecco's medium
- CMP-Neu5Ac hydroxylase:
CMP-N-acetylneuraminate: NAD(P)H oxido-reductase (N-acetyl hydroxylating) (EC 126.96.36.199); CMP-sialate hydrolase (EC 188.8.131.52); sialic acid-pyruvate lyase (EC 184.108.40.206)
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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