Activation of protein kinase C by the dihydropyridine calcium channel blocker, felodipine.

Felodipine, a dihydropyridine Ca2+ channel blocker, appears to have intracellular sites of action in addition to its ability to attenuate voltage-dependent Ca2+ channels in smooth muscle cells. In vitro, felodipine inhibits several calmodulin-dependent enzymes such as myosin light chain kinase, cyclic nucleotide phosphodiesterase and caldesmon kinase [Walsh MP, Sutherland C and Scott-Woo GC, Biochem Pharmacol 37: 1569-1580, 1988]. Such effects may partially explain the relaxant effects of felodipine and related dihydropyridines on vascular smooth muscle. We have examined the effects of felodipine on the activity of another important enzyme which has been implicated in the regulation of the contractile state of smooth muscle, protein kinase C. We chose to use a physiologically relevant substrate of protein kinase C for these studies, viz. platelet P47 protein, rather than the more commonly used lysine-rich histone which is probably not a physiologically important substrate. Protein kinase C and P47 were purified from human platelets and their important structural and functional properties were characterized. Felodipine and the p-chloro analogue of felodipine enhanced both the rate and extent of P47 phosphorylation by protein kinase C. Half-maximal activation was observed at 9.5 microM felodipine and 8.5 microM p-chloro analogue. Activation by felodipine was dependent upon the presence of phospholipid but did not require diacylglycerol. These observations suggest that the pharmacological actions of felodipine and related dihydropyridines may involve activation of protein kinase C in addition to their known effects on voltage-dependent Ca2+ channels and calmodulin-dependent enzymes.