Resolution on major protein kinase substrates in neutrophil cytosol in response to DAG/PS and arachidonic acid stimulation and the selective action of various protein kinase inhibitors

Abstract

Stimulation of human neutrophils induces phosphorylation of several cellular proteins. Human neutrophils possess calcium-dependent protein kinase C (PKC) α and β isoforms and calcium-independent n isoforms. Little is known, however, of the physiological substrates of each isoform. In this study, we characterized the substrates of calcium-dependent and -independent PKC isoforms and the substrate of PKC activated by arachidonic acid. Furthermore, we found that the PKC inhibitor H-7 failed to inhibit phosphorylation of endogenous substrates of calcium-independent PKC activity. These results may help to understand the role of PKC in neutrophil activation and shed light on the different responses elicited by H-7 in intact cells.