Immunological identification of the heart myoglobin radical formed by hydrogen peroxide.
This study reports the detection of protein free radicals using the specific free radical reactivity of nitrone spin traps in conjunction with nitrone-antibody specificity. Polyclonal antibodies were developed that bind to protein adducts of the nitrone spin-trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO). The antibodies were used to detect DMPO protein adducts produced on horse myoglobin resulting from self-peroxidation. Western blot analysis demonstrates that myoglobin forms the predominant radical-derived nitrone adduct in rat heart supernatant.
Laboratory of Pharmacology and Chemistry, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.
Charles D
CD
Detweiler
Leesa J
LJ
Deterding
Kenneth B
KB
Tomer
Colin F
CF
Chignell
Dori
D
Germolec
Ronald P
RP
Mason
D000818:Animals; D000917:Antibody Formation; D015153:Blotting, Western; D003497:Cyclic N-Oxides / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D004797:Enzyme-Linked Immunosorbent Assay; D005609:Free Radicals / Q000378:metabolism; D006321:Heart / Q000187:drug effects*; D006736:Horses; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008786:Metmyoglobin / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D009211:Myoglobin / Q000378:metabolism*; D009589:Nitrogen Oxides / Q000737:chemistry; D010084:Oxidation-Reduction; D011817:Rabbits; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013113:Spin Labels
D003497:Cyclic N-Oxides; D005609:Free Radicals; D009211:Myoglobin; D009589:Nitrogen Oxides; D013113:Spin Labels; C015388:nitrones; D008786:Metmyoglobin; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D006861:Hydrogen Peroxide