DL-threo-beta-fluoroasparagine inhibits asparagine-linked glycosylation in cell-free lysates.

The effect of beta-fluoroasparagine on N-linked glycosylation was examined in a cell-free translation system in which glycosylation is coupled to protein synthesis. The threo-isomer markedly inhibited glycosylation at a concentration of 1 mM, and this effect was blocked by L-asparagine, indicating that glycosylation was inhibited secondary to incorporation of the asparagine analog into protein. The erythro-isomer, at similar concentrations, was not incorporated into protein and had no effect on glycosylation. threo-beta-Fluoroasparagine is highly toxic to some mammalian cells in culture. Our observations suggest that its toxicity may be due in part to the failure of the fluoroasparagine-containing protein to become glycosylated. The data suggest that this analog will be useful for examining the structural determinants for glycosylation.