Does calmodulin play a role in the regulation of cardiac sarcolemmal adenylate cyclase activity?
The recent suggestion that calmodulin (CaM) could mediate calcium inhibition of cardiac adenylate cyclase (AC) has been reassessed. Using a purified sarcolemmal preparation (SL), the influence of different concentrations of free Ca2+ (obtained using Ca2+-EGTA solutions) was studied on dog heart AC. From 10(-9) M to 10(-3) M Ca2+ reduced basal activity, as well as epinephrine (10(-4) M)- and trypsin (1.0 microgram/mL)-stimulated activities with, in the three cases, an identical IC50 of 10(-8) M. The amount of endogenous CaM in the SL, measured using a radioimmunoassay technique, was found to be 7.5 ng/mg protein. The resulting concentration of CaM in the final AC incubation medium was lower than 50 pM, indicating the lack of a significant role for endogenous CaM in the inhibition observed. The addition of exogenous CaM to the AC assay at a concentration sufficient to stimulate other CaM-dependent systems did not modify the Ca2+ inhibitory curves for basal, epinephrine (10(-4) M)-stimula
