Detection of secondary radicals from peroxynitrite-mediated oxidations by electron spin resonance

Methods in Enzymology

Volumn 269, Issue , 1996, Pages 346-354

  Augusto, Ohara ^a   Radi, Rafael ^a   Gatti, Reynaldo M ^a   Vásquez Vivar, Jeannette ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

FREE RADICAL; NITRIC OXIDE; NITRITE; PEROXYACETYLNITRATE; SUPEROXIDE;

ARTICLE; CYTOTOXICITY; ELECTRON SPIN RESONANCE; OXIDATIVE STRESS; PRIORITY JOURNAL;

EID: 0029792288     PISSN: 00766879     EISSN: None     Source Type: Book Series    
DOI: 10.1016/s0076-6879(96)69035-1     Document Type: Article

References (21)

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16. 15 This has been confirmed by systematic studies with ultrapure DMPO that demonstrated that preformed DMPO-hydroxyl radical adduct (3-4 μM) is destroyed in a concentration-dependent manner by peroxynitrite, with 20 μM oxidant being required to destroy 90% of the adduct at pH 6.0 and 7.5. In the presence of 20 μM GSH, only 30% of the DMPO-hydroxyl radical adduct is destroyed by 20 μM oxidant, confirming the protective effect of thiol groups on peroxynitrite-mediated adduct decay. Similar results were obtained with preformed PBN-methyl radical adduct, which at 0.4 μM concentration was completely destroyed by 10 μM peroxynitrite; this adduct was also protected by GSH. Peroxynitrite, however, was unable to destroy nitroxides that lack hydrogens in the α-carbon, such as 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL). Similarly, carbon-centered radical adducts of the spin trap DBNBS (3,5-dibromo-4-nitrosobenzenesulfonic acid) appear to be resistant to peroxynitrite because a strong DBNBS-methyl radical adduct signal is observed during peroxynitrite-mediated oxidation of dimethyl sulfoxide in the absence of thiols.
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19. 15,16 Recently, we have also observed that in the absence of oxidizable substrates, peroxynitrite is able to nitrate the spin trap PBN.
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