Immuno-spin trapping detection of protein-centered radicals

Producción CyT

Artículo

Autoría

RAMIREZ, DARIO

Fecha

2005

Editorial y Lugar de Edición

JOHN WILEY & SONS INC

Revista

Current Protocols in Toxicology, vol. 17 (pp. 1-23) - ISSN 1934-9254
JOHN WILEY & SONS INC

ISSN

1934-9254

Resumen Información suministrada por el agente en SIGEVA

Protein-centered radicals are involved in biological oxidative damage induced by drugs, environmental hazards, and cellular reactive oxygen species. Presently, the technique most widely used to study protein-centered radicals is electron spin resonance (ESR; also known as electron paramagnetic resonance, EPR); used either directly or in combination with the spin-trapping technique. Protein-centered radicals may be trapped with the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) formin... Protein-centered radicals are involved in biological oxidative damage induced by drugs, environmental hazards, and cellular reactive oxygen species. Presently, the technique most widely used to study protein-centered radicals is electron spin resonance (ESR; also known as electron paramagnetic resonance, EPR); used either directly or in combination with the spin-trapping technique. Protein-centered radicals may be trapped with the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) forming DMPO-radical adducts. However, after a few minutes these adducts decay, often by oxidation to DMPO-protein radical–derived nitrone adducts, which are ESR-silent species. Because nitrone adducts are not free radicals and their formation involves the creation of a covalent linkage, they are stable long after the ESR signal decays. In the new alternative technique of immuno-spin trapping, nitrone adducts are detected by using an antibody, i.e., anti-DMPO, that recognizes their nitrone moiety. Immuno-spin trapping is a simple, reliable, affordable, sensitive, and specific approach to detecting protein-centered radicals, and its development brings the power of immunoassays to bear on the field of toxicology of free radical–mediated biological damage.

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Palabras Clave

OXIDATIVE STRESNITRONE ADDUCTOXIDATIVE STRESSPROTEIN OXIDATION