Use of a genetic variant to study the hexose transport properties of human skin fibroblasts.
Human skin fibroblasts from 'normal' subjects were found to possess at least two hexose transport systems. One system was responsible for the uptake of 2-deoxy-D-glucose (dGlc), D-glucose and D-galactose, whereas the other was responsible primarily for the uptake of 3-O-methyl-D-glucose (MeGlc). The transport of dGlc was the rate-limiting step in the uptake process; over 97% of the internalized dGlc was phosphorylated and the specific activity of hexokinase was several times higher than that for dGlc transport. The dGlc transport system was activated by glucose starvation, and was very sensitive to inhibition by cytochalasin B and energy uncouplers. Fibroblasts isolated from a patient with symptoms of hypoglycaemia were found to differ from their normal counterparts in the dGlc transport system. They exhibited a much higher transport affinity for dGlc, D-glucose and D-galactose, with no change in the respective transport capacity. Transport was not the rate-limiting step in dGlc uptake by these cells. Moreover, the patient's dGlc transport system was no longer sensitive to inhibition by cytochalasin B and energy uncouplers. This suggested that the intrinsic properties of the patient's dGlc transport system were altered. It should be noted that the patient's dGlc transport system could still be activated by glucose starvation. Despite the changes in the dGlc transport system, the MeGlc transport system in the patient's fibroblasts remained unaltered. The observed difference in the properties of the two hexose transport systems in the 'normal' and the patient's fibroblasts strongly suggests that the two transport systems may be coded or regulated by different genes. The present finding provides the first genetic evidence from naturally occurring fibroblasts indicating the presence of two different hexose transport systems.
Mesmer
O T
OT
Department of Biochemistry, University of Western Ontario, London, Canada.
Gordon
B A
BA
Rupar
C A
CA
Lo
T C
TC
D019325:3-O-Methylglucose; D001692:Biological Transport; D003571:Cytochalasin B / Q000494:pharmacology; D003847:Deoxyglucose / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006601:Hexoses / Q000378:metabolism*; D006801:Humans; D007003:Hypoglycemia / Q000235:genetics; D007700:Kinetics; D008757:Methylglucosides / Q000378:metabolism; D012867:Skin / Q000378:metabolism*; D014475:Uncoupling Agents
D006601:Hexoses; D008757:Methylglucosides; D014475:Uncoupling Agents; D019325:3-O-Methylglucose; D003571:Cytochalasin B; D003847:Deoxyglucose; D005947:Glucose; D005690:Galactose