Document Detail


EPR detection of glutathionyl and protein-tyrosyl radicals during the interaction of peroxynitrite with macrophages (J774).
MedLine Citation:
PMID:  11518708     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Peroxynitrite is one of the biological oxidants whose addition to cells has been shown to either activate signaling pathways or lead to cell injury, depending on cell type and oxidant concentration. The intermediacy of free radicals in these processes has been directly demonstrated only during the interaction of peroxynitrite with erythrocytes, a particular cell type, due to its high hemoglobin content. Here, we demonstrate that the addition of peroxynitrite to a macrophage cell line (J774) led to the production of glutathionyl and protein-tyrosyl radicals. The glutathionyl radical was characterized by EPR spin-trapping experiments with 5,5-dimethyl-1-pyrroline-N-oxide. Protein-tyrosyl radical formation was suggested by direct EPR spectroscopy and confirmed by EPR spin-trapping experiments with 3,5-dibromo-4-nitrosobenzenesulfonic acid and Western blot analysis of nitrated proteins in treated macrophages. Time dependence studies of free radical formation indicate that intracellular glutathione and unidentified proteins are the initial peroxynitrite targets in macrophages and that their derived radicals trigger radical chain reactions. The results are likely to be relevant to the understanding of the bioregulatory and biodamaging effects of peroxynitrite.
Authors:
S Lopes de Menezes; O Augusto
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2001-08-22
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  276     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2001 Oct 
Date Detail:
Created Date:  2001-10-22     Completed Date:  2001-12-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  39879-84     Citation Subset:  IM    
Affiliation:
Departamento de Bioquimica, Instituto de Quimica, Universidade de São Paulo, CP 26077, CEP 05513-970, São Paulo, SP, Brazil.
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MeSH Terms
Descriptor/Qualifier:
Animals
Benzenesulfonates
Cell Survival
Cells, Cultured
Cyclic N-Oxides
Electron Spin Resonance Spectroscopy
Free Radicals
Glutathione / analysis*
Macrophages / cytology,  metabolism*
Mice
Models, Chemical
Nitroso Compounds
Peroxynitrous Acid / metabolism*
Proteins / analysis*
Spin Labels
Sulfhydryl Compounds / metabolism
Tyrosine
Chemical
Reg. No./Substance:
0/Benzenesulfonates; 0/Cyclic N-Oxides; 0/Free Radicals; 0/Nitroso Compounds; 0/Proteins; 0/Spin Labels; 0/Sulfhydryl Compounds; 14691-52-2/Peroxynitrous Acid; 3317-61-1/5,5-dimethyl-1-pyrroline-1-oxide; 55520-40-6/Tyrosine; 70-18-8/Glutathione; 83016-63-1/3,5-dibromo-4-nitrosobenzenesulfonate

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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