Effect of inhibitors of N-linked oligosaccharide processing on the high-affinity transport of D-aspartate by C6 glioma cells.

The effect of several inhibitors of oligosaccharide-processing on the high-affinity transport of D-aspartate was investigated in C6 glioma cells. Swainsonine, an inhibitor of mannosidase II, had no effect on the uptake of the amino acid. Castanospermine (100 micrograms/ml) and 1-deoxynojirimycin (1 mM), inhibitors of glucosidases, and 1-deoxymannojirimycin (1 mM), an inhibitor of mannosidase I, reduced the rate of transport by 35-45%. All inhibitory compounds decreased the Vmax for transport without affecting the Km which suggests that inhibition of oligosaccharide trimming reduces the number of competent transporters on the surface of the plasma membrane. Returning the cells to a drug-free medium for 24 h, following a 24 h exposure, resulted in complete recovery of uptake. Treatment of cells with neuraminidase from V. cholerae also decreased the Vmax for transport by about 20%. The results suggest that: (i) a partial complex carbohydrate chain on the high-affinity transporter for acidic amino acid transmitters is sufficient for activity and (ii) sialic acid residues may be necessary for normal operation of the transporter.