Coupling of insulin binding and insulin action on glucose transport in fat cells

The initial lag phase of insulin action on glucose transport in adipocytes reflects an unknown process that couples receptor binding and glucose transport activity. The influence of temperature, cellular ATP, cyclic AMP, and calcium on this process and a possible relation to internalization of insulin were studied. The Arrhenius plot of the coupling shows a break in slope at 30 degrees C; the activation energy below 30 degrees C is 17.5 kcal/mol. Reduction of cellular ATP by 70% prolongs the coupling process; initial binding and final maximal response of the glucose transport remain unaffected. Further reduction of ATP (greater than 90%) before addition of insulin abolishes the coupling completely. Reduction of ATP at different time points after addition of insulin blocks further activation; however, the actual state of activity is preserved. Calcium depletion by EDTA prolongs the coupling and decreases the maximal response. Internalization of insulin as determined in chloroquine-treated cells begins later than transport activation and is in contrast to transport activation not observable at 15 degrees C. In conclusion, the coupling is not related to internalization; it is ATP-dependent, whereas the initial binding and the activated transport system are ATP-independent. Calcium but not cyclic AMP might be second messenger or cofactor in the coupling process.