The role of C-kinase in the activation of human polymorphonuclear leukocytes has been examined using H-7, a recently described C-kinase inhibitor. We found that H-7 will inhibit PMN superoxide anion release in response to the tumor promotor phorbol myristate acetate and the calcium ionophore A23187. In contrast, no inhibition by H-7 was seen for PMN superoxide release stimulated by the chemotactic peptide FMLP. H-7 did not inhibit PMN NADPH oxidase activity or PMN degranulation by any stimulant, but it reversed a phorbol ester-induced inhibition of granule release by FMLP. The results show that H-7 differentially affects the PMN functional events of secretion and superoxide release and suggests that an H-7 inhibitable C-kinase is not involved in chemotactic peptide induced activation of PMN and may not regulate stimulus induced PMN degranulation.