Neuraminidase treatment of isolated rat adipocytes and differential regulation of basal and insulin-stimulated glucose transport.
The role of membrane carbohydrate in the function of insulin receptors and glucose transporters was investigated with neuraminidase to release sialic acid from isolated rat adipocytes. Pretreatment of adipocytes with neuraminidase resulted in an increase in basal glucose transport. At the same time, insulin-stimulated glucose transport was reduced by an average of 30%. The enzyme action on glucose transport was not due to a nonspecific membrane perturbation because neuraminidase caused only a nonsignificant decrease in the uptake of the amino acid analog alpha-(methylamino)isobutyric acid and had no effect on basal or insulin-stimulated protein synthesis. Insulin binding was slightly increased in neuraminidase-treated cells, yet the shapes of the dose-response curves for insulin stimulation of glucose transport were similar (EC50 = 0.087 +/- 0.010 and 0.082 +/- 0.008 nM for control and treated cells, respectively). The neuraminidase-induced increase in basal transport was the result of an increase in the affinity of the glucose-transport system (Km = 7.3 +/- 1.4 and 3.6 +/- 0.7 mM before and after treatment, respectively) with no change in Vmax. Conversely, enzyme treatment decreased the Vmax of insulin-stimulated 3-O-methylglucose transport from 87.8 +/- 13.2 to 41.3 +/- 4.9 pmol.2 x 10(5) cells-1.s-1 while not altering the Km. These changes in glucose-transport activity resulting from enzyme treatment were not due to alterations in glucose-transporter concentration on the plasma membrane as measured by the D-glucose-inhibitable binding of [3H]cytochalasin B. Thus, sialic acid plays multiple roles in the control of glucose-transport activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Ciaraldi
T P
TP
Veterans Administration Medical Center, Medical Research Service, San Diego, La Jolla, California.
D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D002469:Cell Separation; D004305:Dose-Response Relationship, Drug; D005947:Glucose / Q000378:metabolism / Q000493:pharmacokinetics*; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology*; D009004:Monosaccharide Transport Proteins / Q000378:metabolism / Q000502:physiology; D009439:Neuraminidase / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D011972:Receptor, Insulin / Q000378:metabolism / Q000502:physiology; D012794:Sialic Acids / Q000032:analysis / Q000502:physiology
D007328:Insulin; D009004:Monosaccharide Transport Proteins; D012794:Sialic Acids; D011972:Receptor, Insulin; D009439:Neuraminidase; D005947:Glucose