Membrane endopeptidases of human neutrophil.

Recent studies using inhibitors or synthetic substrates of serine protease suggest that membrane protease activity may be essential for neutrophil chemotaxis, phagocytosis, degranulation, and superoxide production. However, little is known about the nature and localization of the proteases. In this study, we demonstrated that intact human neutrophils hydrolyzed [125I]glucagon. The degradation of glucagon was temperature dependent and was not dependent on the release of lysosomal enzymes. Two endopeptidases were demonstrated: a metalloendopeptidase which accounted for two thirds of the intact cell activity, and a serine endopeptidase, accounting for the rest of the activity. Both enzymes had a neutral to alkaline pH optimum (pH 7-9). The metalloendopeptidase had a Km of 15.3 microM and Vmax of 5.9 nmol/5 X 10(6) cells/45 min. The corresponding values for the serine endopeptidase were 33.3 microM and 5.0 nmol/5 X 10(6) cells/45 min, respectively. Inhibition of the membrane metalloendopeptidase or serine endopeptidase by 1,10-phenanthroline or diisopropylfluorophosphate, respectively, did not inhibit the production of superoxide by phorbol myristate acetate-stimulated neutrophils.