Increased hexose transport in Chinese hamster ovary cells resistant to 3-O-methyl-D-glucose

3-O-Methyl-D-glucose-resistant mutants were selected from Chinese hamster ovary cells after mutagenesis with ethyl methanesulfonate. A mutant, MegR24, was isolated which was significantly more resistant than the parent to 3-O-methylglucose. Uptake of 50 microM D-glucose by metabolizing MegR24 cells was 2- to 3-fold higher than the parental cells in the absence or presence of 100 mM 3-O-methylglucose. A study of transport of D-[3H]glucose in ATP-depleted cells indicated an apparent Km for D-glucose transport that was 3-fold lower for the mutant (2.7 +/- 0.3 mM) than for the parent (8.9 +/- 1.0 mM). The apparent Km for transport of 3-O-methylglucose by the mutant (10.9 +/- 2.4 mM) was almost 2-fold lower than that of the parent (21.4 +/- 4.7 mM). The Vmax values for transport of D-glucose or 3-O-methylglucose by the mutant and parental cell lines were not significantly different. The MegR24 mutant also exhibited enhanced countertransport of D-[3H]glucose following preloading of ATP-depleted cells with 100 mM 3-O-methylglucose. Simple diffusion of hexoses as measured by L-glucose uptake was not altered in the mutant. These results suggest that the MegR24 hexose carrier has an increased affinity for transport of hexoses and that the resistance to the cytotoxic effects of 3-O-methylglucose exhibited by MegR24 is due to its ability to transport D-glucose 2- to 3-fold more efficiently than the parental strain.