Effects of divalent metals on the specificity of inhibitors of the cyclic nucleotide phosphodiesterases from bovine heart

Abstract

Divalent metals used to support phosphodiesterase (EC 3.1.4.-) activity have been found to influence the substrate and enzyme specificity of many phosphodiesterase inhibitors in studies of the hydrolysis of cyclic AMP and cyclic GMP by the calmodulin-dependent and cyclic AMP-specific phosphodiesterases from bovine heart. Many compounds displayed marked differences in substrate specificity and inhibitory potency in the presence of Mg 2+, as compared with Mn 2+, when studied with the unactivated form of calmodulin-dependent phosphodiesterase, while few compounds displayed differences in the presence of calmodulin. With a single divalent metal, marked differences in inhibitory potency and substrate specificity were also observed in the absence or presence of calmodulin suggesting that alterations in calmodulin and/or Ca 2+ levels may greatly affect the response to phosphodiesterase inhibitors. Divalent metals did not alter the effects of inhibitors on the hydrolysis of cyclic AMP by the cyclic AMP-specific phosphodiesterase, however divalent metals would probably indirectly influence the relative cellular level of cyclic AMP hydrolyzed by this enzyme, and therefore the effects of inhibitors, through metal effects on the calmodulin-dependent phosphodiesterase. No correlation was found between the inhibitory activity of the compounds, many of which were cyclic nucleotide analogs, and their ability to activate cyclic AMP-dependent or cyclic GMP-dependent protein kinases or to affect cyclic AMP-dependent protein kinase activity by displacing bound cyclic AMP.