Suitability of 2-deoxyglucose for measuring initial rates of glucose uptake in isolated adipocytes.
The suitability of [3H]-2-deoxyglucose from measuring initial rates of glucose uptake in isolated rat adipocytes was assessed using three approaches. Basal and insulin-stimulated rates of glucose uptake were directly compared in 2 sec and 5 min assays using [14C]-3-O-methylglucose, [3H]-2-deoxyglucose, and [3H]-D-glucose. Equilibrium kinetics of 2-deoxyglucose uptake were compared with those of 3-O-methylglucose through impairment of hexokinase activity by depleting cellular energy with 2,4-dinitrophenol. The equivalence of these glucose analogues in a dynamic system was assessed by measuring the lag time preceding insulin stimulation of glucose uptake, insulin activation rates, and the T 1/2 of insulin activation. Our results demonstrate that no fundamental difference exists in the initial transport of 3-O-methylglucose, 2-deoxyglucose, and D-glucose.
Traxinger
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Department of Biochemistry, University of Tennessee, Memphis 38163.
Marshall
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S
D019297:2,4-Dinitrophenol; D019325:3-O-Methylglucose; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D003847:Deoxyglucose / Q000378:metabolism*; D004140:Dinitrophenols / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism*; D006593:Hexokinase / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008757:Methylglucosides / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D004140:Dinitrophenols; D007328:Insulin; D008757:Methylglucosides; D019325:3-O-Methylglucose; D003847:Deoxyglucose; D006593:Hexokinase; D005947:Glucose; D019297:2,4-Dinitrophenol