title: Inactivation of enzymes and oxidative modification of proteins by stimulated neutrophils.
abstract: Differentiated, stimulated HL-60 cells and freshly isolated, stimulated neutrophils inactivate glutamine synthetase (L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2) either inside or outside of Escherichia coli. Stimulated neutrophils also inactivate at least four endogenous enzymes which are inactivated by mixed-function oxidation (MFO) systems in vitro (L. Fucci, C.N. Oliver, M.J. Coon, and E.R. Stadtman (1983) Proc. Natl. Acad. Sci. USA 80, 1521-1525). The inactivation of glutamine synthetase by stimulated neutrophils exhibits characteristics similar to those previously described using both enzymic and nonenzymic MFO systems (R.L. Levine, C.N. Oliver, R.M. Fulks, and E.R. Stadtman (1981) Proc. Natl. Acad. Sci. USA 78, 2120-2124). Although the reaction occurs in the absence of Fe(III), it is stimulated by added Fe (III). Inactivation required molecular oxygen and is partially inhibited by Mn(II), catalase, superoxide dismutase, and metal chelators, ethylenediaminetetraacetic acid and o-phenanthroline. Both the kinetics and the extent of glutamine synthetase inactivation differ when neutrophils are stimulated with phorbol esters compared with formylated peptides. Glutamine synthetase inactivation catalyzed by MFO systems is accompanied by the formation of protein carbonyl derivatives which form stable hydrazones when treated with 2,4-dinitrophenylhydrazine. Multiple carbonyl derivatives are formed in the soluble protein fraction of stimulated neutrophils and these derivatives collectively exhibit an absorbance spectrum similar to that of glutamine synthetase inactivated by liver microsomal cytochrome P-450 MFO system (K. Nakamura, C.N. Oliver, and E.R. Stadtman (1985) Arch. Biochem. Biophys. 240, 319-329).
mesh_terms: D001770:Blood Bactericidal Activity; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D055598:Chemical Phenomena; D002621:Chemistry; D005290:Ferric Compounds / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000037:antagonists & inhibitors*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D002614:Chelating Agents; D005290:Ferric Compounds; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D006861:Hydrogen Peroxide; D005974:Glutamate-Ammonia Ligase; D013755:Tetradecanoylphorbol Acetate