Immuno-spin trapping of protein and DNA radicals: "tagging" free radicals to locate and understand the redox process.
Biomolecule-centered radicals are intermediate species produced during both reversible (redox modulation) and irreversible (oxidative stress) oxidative modification of biomolecules. These oxidative processes must be studied in situ and in real time to understand the molecular mechanism of cell adaptation or death in response to changes in the extracellular environment. In this regard, we have developed and validated immuno-spin trapping to tag the redox process, tracing the oxidatively generated modification of biomolecules, in situ and in real time, by detecting protein- and DNA-centered radicals. The purpose of this methods article is to introduce and update the basic methods and applications of immuno-spin trapping for the study of redox biochemistry in oxidative stress and redox regulation. We describe in detail the production, detection, and location of protein and DNA radicals in biochemical systems, cells, and tissues, and in the whole animal as well, by using immuno-spin trapping with the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide.
Gomez-Mejiba
Sandra E
SE
Free Radical Biology and Aging Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA.
Zhai
Zili
Z
Akram
Hammad
H
Deterding
Leesa J
LJ
Hensley
Kenneth
K
Smith
Nataliya
N
Towner
Rheal A
RA
Tomer
Kenneth B
KB
Mason
Ronald P
RP
Ramirez
Dario C
DC
D000818:Animals; D001671:Biochemistry / Q000379:methods; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D035843:Biomedical Research; D002460:Cell Line; D003497:Cyclic N-Oxides / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000032:analysis; D005609:Free Radicals / Q000032:analysis; D008264:Macrophages / Q000737:chemistry / Q000378:metabolism*; D008279:Magnetic Resonance Imaging; D013058:Mass Spectrometry; D051379:Mice; D008956:Models, Chemical; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011485:Protein Binding; D012680:Sensitivity and Specificity; D019097:Spin Trapping / Q000295:instrumentation / Q000379:methods*
D015415:Biomarkers; D003497:Cyclic N-Oxides; D005609:Free Radicals; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D004247:DNA