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J. Biol. Chem., Vol. 260, Issue 5, 2646-2652, Mar, 1985
SC Frost and MD Lane
Following the differentiation of 3T3-L1 preadipocytes insulin acutely
activates the rate of 2-deoxy-[1-14C]glucose uptake in the mature 3T3- L1
adipocyte by 15- to 20-fold. Phenylarsine oxide, a trivalent arsenical that
forms stable ring complexes with vicinal dithiols, prevents
insulin-activated hexose uptake in a concentration-dependent manner (Ki = 7
microM) but has no inhibitory effect on basal hexose uptake.
2,3-Dimercaptopropanol at a level nearly stoichiometric to that of
phenylarsine oxide prevents or rapidly reverses the inhibition of hexose
uptake; 2-mercaptoethanol, even in high stoichiometric excess over the
arsenical, does not reverse inhibition of hexose uptake. When phenylarsine
oxide is added after adipocytes have been fully activated by insulin,
2-deoxy-[1-14C]glucose uptake rate decays slowly at a rate corresponding to
that caused by the withdrawal of insulin (t1/2 = 10 min). Using the same
conditions under which phenylarsine oxide blocked activation, the Km for
deoxyglucose uptake, the rate at which 125I- insulin became
cell-associated, and the 125I-insulin binding isotherm for solubilized
insulin receptor were not affected by phenylarsine oxide. These results
support the transporter translocation model for insulin-activated hexose
transport and implicate vicinal sulfhydryl groups in a post-insulin binding
event essential for the translocation of glucose transporters to the plasma
membrane.
Evidence for the involvement of vicinal sulfhydryl groups in insulin- activated hexose transport by 3T3-L1 adipocytes
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