title: Protein nitration is predominantly mediated by a peroxynitrite-dependent pathway in cultured human leucocytes.
abstract: Protein nitration is a common characteristic of oxidative injury caused by the invasion of leucocytes into inflammatory lesions. Two distinct pathways of nitration of protein tyrosine residues, namely the peroxynitrite (ONOO(-))-mediated pathway and another catalysed by the haem-containing peroxidases, have been reported under experimental conditions. However, the contribution of these two pathways in human leucocytes is still controversial. The present study demonstrates that the process of phenolic nitration of proteins in cultured human leucocytes is mainly ONOO(-)-mediated and that it differs between granulocytes and mononuclear cells, depending on the cell compartment and the stimuli. We have also shown that NO induces protein nitration via a ONOO(-)-dependent pathway, whereas NO(2)(-), the NO metabolite, does not increase but decreases nitration in PMA-stimulated leucocytes. The inhibition of myeloperoxidase activity did not reduce protein nitration; on the other hand, the myeloperoxidase inhibitor aminobenzoic hydrazide caused increased nitration, which was mediated by ONOO(-). These results suggest that protein nitration is predominantly mediated by a ONOO(-)-dependent pathway in cultured human leucocytes and that the myeloperoxidase-catalysed pathway does not play a significant role in protein nitration.
authors.affiliation: Department of Integrative Human Cardiovascular Physiology and Functional Genomics, Division of Cardiac Surgery, University of Leicester, Glenfield Hospital, Leicester LE3 9QP, U.K. mg50@le.ac.uk
authors.forename: Manuel
authors.initials: M
authors.lastname: Galiñanes
authors.forename: Bashir M
authors.initials: BM
authors.lastname: Matata
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology; D002478:Cells, Cultured; D005110:Extracellular Space / Q000378:metabolism; D006801:Humans; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism*; D008665:Metalloporphyrins / Q000494:pharmacology; D009566:Nitrates / Q000378:metabolism; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D013096:Spermine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: C466420:5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrinato iron(III) chloride; D000814:Aniline Compounds; D008665:Metalloporphyrins; D009566:Nitrates; D011506:Proteins; D030421:Peroxynitrous Acid; D013096:Spermine; C002744:3-nitrotyrosine; D014443:Tyrosine; C034343:4-aminobenzhydrazide; D009195:Peroxidase; D013755:Tetradecanoylphorbol Acetate