Energy-dependent and protein synthesis-independent recycling of the insulin-sensitive glucose transport mechanism in fat cells.
The glucose transport mechanism of rat epididymal fat cells was reconstituted into egg lecithin liposomes, and their carrier-mediated transport activity ws estimated from the difference in the rates of uptake of D-[3H]glucose and L-[14C]glucose. Insulin increased the glucose transport activity in the plasma membrane-rich fraction while decreasing the activity in the Golgi-rich fraction in agreement with our previous data (Suzuki, K., and Kono, T. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 2542-2545). The development of the insulin effects was inhibited when cells were exposed to 2,4-dinitrophenol or KCN before the insulin treatment. In addition, the reversal of the insulin effects was blocked upon exposure of insulin-treated cells to 2,4-dinitrophenol or KCN prior to the elimination of the hormone. In contrast, neither development nor reversal of the insulin effects was affected by cycloheximide or puromycin. The temperature coefficients of the transport activities reconstituted from the basal or insulin-treated forms of the plasma membrane-rich or Golgi-rich fractions were all identical. The recoveries of protein, 5'-nucleotidase, UDP-galactose:N-acetylglucosamine galactosyltransferase, and NADH dehydrogenase into subcellular fractions were determined. However, net effects of insulin on the glucose transport activities have remained unknown for lack of an appropriate marker enzyme of the Golgi-like vesicles associated with the transport activity. It is suggested that the glucose transport mechanism is recycled between the plasma membrane-rich and Golgi-rich fractions by an energy-dependent reaction.
D019297:2,4-Dinitrophenol; D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D004140:Dinitrophenols / Q000494:pharmacology; D004734:Energy Metabolism / Q000187:drug effects; D005947:Glucose / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism; D008297:Male; D011190:Potassium Cyanide / Q000494:pharmacology; D051381:Rats
D004140:Dinitrophenols; D007328:Insulin; D008081:Liposomes; D005947:Glucose; D011190:Potassium Cyanide; D019297:2,4-Dinitrophenol