Acceleration of the adipocyte adenylate cyclase turn-off reaction by inhibitory hormonal factors.

beta-Adrenergic agonists such as isoproterenol stimulate hamster adipocyte adenylate cyclase by a GTP-dependent process, whereas prostaglandin E1, alpha-adrenergic agonists, and nicotinic acid inhibit the enzyme by a process also dependent on GTP and amplified by sodium ions. We have determined the first-order rate constant describing the decay of isoproterenol plus GTP-stimulated adenylate cyclase and the modulation of this off rate constant by sodium and inhibitory hormonal factors. With 1 microM GTP and 0.2 mM isoproterenol, the off rate constant was 5.0 min-1 at 25 degrees C. Addition of NaCl (100 mM), which increased basal and isoproterenol-stimulated cyclase activities, decreased the rate constant of the hormone-stimulated enzyme to 1.4 min-1. Prostaglandin E1 (10 microM) and nicotinic acid (30 microM), which decreased basal and hormonally stimulated enzyme activities, increased the NaCl-suppressed off rate constant to 6.1 and 6.0 min-1, respectively. Similar data were obtained with 1 mM isoproterenol with MnATP and MgATP as the adenylate cyclase substrate. On the other hand, the turn-on reaction of adipocyte adenylate cyclase, measured with the stable GTP analogue 5'-guanylyl imidodiphosphate (30 microM), was accelerated by isoproterenol (1 mM) and NaCl (100 mM). Under all of these conditions, inhibitory hormonal agents did not cause any delay in the turn-on reaction. These data indicate that, in hamster adipocyte membranes, inhibitory hormonal factors inhibit adenylate cyclase by increasing the enzyme's turn-off reaction.