TY - JOUR
T1 - Glutathione propagates oxidative stress triggered by myeloperoxidase in HL-60 cells
T2 - Evidence for glutathionyl radical-induced peroxidation of phospholipids and cytotoxicity
AU - Borisenko, Grigory G.
AU - Martin, Ian
AU - Zhao, Qing
AU - Amoseato, Andrew A.
AU - Tyurina, Yulia Y.
AU - Kagan, Valerian E.
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2004/5/28
Y1 - 2004/5/28
N2 - Glutathione acts as a universal scavenger of free radicals at the expense of the formation of the glutathionyl radicals (GS.). Here we demonstrated that GS. radicals specifically interact with a reporter molecule, paramagnetic and non-fluorescent 4-((9-acridinecarbonyl)-amino)-2,2,6,6-tetrametliylpiperidine-1-oxyl (Ac-Tempo), and convert it into a non-paramagnetic fluorescent product, identified as 4-((9-acridinecarbonyl)amino)-2,2,6,6-tetramethylpiperidine (Ac-piperidine). Horseradish peroxidase-, myeloperoxidase-, and cyclooxygenase-catalyzed oxidation of phenol in the presence of H2O2 and GSH caused the generation of phenoxyl radicals and GS. radicals, of which only the latter reacted with Ac-Tempo. Oxidation of several other phenolic compounds (e.g. etoposide and tyrosine) was accompanied by the formation of GS. radicals along with a characteristic fluorescence response from Ac-Tempo. In myeloperoxidase-rich HL-60 cells treated with H 2O2 and phenol, fluorescence microscopic imaging of Ac-Tempo revealed the production of GS. radicals. A thiol-blocking reagent, N-ethylmaleimide, as well as myeloperoxidase inhibitors (succinyl acetone and azide), blocked formation of fluorescent acridine-piperidine. H 2O2/phenol-induced peroxidation of major classes of phospholipids in HL-60 cells was completely inhibited by Ac-Tempo, indicating that GS. radicals were responsible for phospholipid peroxidation. Thus, GSH, commonly viewed as a universal free radical scavenger and major intracellular antioxidant, acts as a pro-oxidant during myeloperoxidase-catalyzed metabolism of phenol in HL-60 cells.
AB - Glutathione acts as a universal scavenger of free radicals at the expense of the formation of the glutathionyl radicals (GS.). Here we demonstrated that GS. radicals specifically interact with a reporter molecule, paramagnetic and non-fluorescent 4-((9-acridinecarbonyl)-amino)-2,2,6,6-tetrametliylpiperidine-1-oxyl (Ac-Tempo), and convert it into a non-paramagnetic fluorescent product, identified as 4-((9-acridinecarbonyl)amino)-2,2,6,6-tetramethylpiperidine (Ac-piperidine). Horseradish peroxidase-, myeloperoxidase-, and cyclooxygenase-catalyzed oxidation of phenol in the presence of H2O2 and GSH caused the generation of phenoxyl radicals and GS. radicals, of which only the latter reacted with Ac-Tempo. Oxidation of several other phenolic compounds (e.g. etoposide and tyrosine) was accompanied by the formation of GS. radicals along with a characteristic fluorescence response from Ac-Tempo. In myeloperoxidase-rich HL-60 cells treated with H 2O2 and phenol, fluorescence microscopic imaging of Ac-Tempo revealed the production of GS. radicals. A thiol-blocking reagent, N-ethylmaleimide, as well as myeloperoxidase inhibitors (succinyl acetone and azide), blocked formation of fluorescent acridine-piperidine. H 2O2/phenol-induced peroxidation of major classes of phospholipids in HL-60 cells was completely inhibited by Ac-Tempo, indicating that GS. radicals were responsible for phospholipid peroxidation. Thus, GSH, commonly viewed as a universal free radical scavenger and major intracellular antioxidant, acts as a pro-oxidant during myeloperoxidase-catalyzed metabolism of phenol in HL-60 cells.
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U2 - 10.1074/jbc.M400119200
DO - 10.1074/jbc.M400119200
M3 - Article
C2 - 15039448
AN - SCOPUS:2542428557
SN - 0021-9258
VL - 279
SP - 23453
EP - 23462
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 22
ER -