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EPR spin-trapping of protein radicals to investigate biological oxidative mechanisms

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Summary.

Presently, free radicals and oxidants are considered to mediate from signaling circuits involved in physiology and pathology to cell and tissue injury. The elucidation of these many inter-related processes requires a better understanding of cellular oxidative mechanisms many of which are mediated by protein radicals. Here, we will discuss the potentialities of EPR spin-trapping of protein radicals to unravel oxidative mechanisms. An overview of the methodology and its application to identify protein residues that are the target of specific oxidants, characterize emerging oxidants, and discriminate radical from non radical mechanisms will be presented. The examples are based on work developed in our laboratories but will be discussed in a broad scenario to emphasize that simple experiments can provide relevant insights into the biological reactivity of known and emerging biological oxidants and into signaling mechanisms.

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Abbreviations

BSA:

bovine serum albumin

Cu,Zn-SOD:

superoxide dismutase

DBNBS:

3,5-dibromo-4-nitrosobenzenesulfonic acid

DMPO:

5,5-dimethylpyrroline N-oxide

EPR:

electron paramagnetic resonance

ESI:

electrospray ionization

GSH:

glutathione

Hb:

hemoglobin

HCO4 − :

peroxycarbonate or peroxymonocarbonate

HSA:

human serum albumin

LC:

liquid chromatography

MALDI-TOF:

matrix-assisted laser desorption/ionization time of flight

MNP:

2-methyl-2-nitrosopropane

MS:

mass spectrometry

PBN:

phenyl-N-t-butylnitrone

Peroxynitrite:

sum of peroxynitrite anion (ONOO−, oxoperoxonitrate (−1)) and peroxynitrous acid (ONOOH, hydrogen oxoperoxonitrate) unless specified

RNS:

reactive nitrogen species

ROS:

reactive oxygen species

References

  • H Arai BS Berlett PB Chock ER Stadtman (2005) ArticleTitleEffect of bicarbonate on iron-mediated oxidation of low-density lipoprotein Proc Natl Acad Sci USA 102 10472–10477 Occurrence Handle16027354 Occurrence Handle10.1073/pnas.0504685102 Occurrence Handle1:CAS:528:DC%2BD2MXntVSitL4%3D

    Article  PubMed  CAS  Google Scholar 

  • F Aslund J Beckwith (1999) ArticleTitleBridge over troubled waters: sensing stress by dissulfide bond formation Cell 96 751–753 Occurrence Handle10102262 Occurrence Handle10.1016/S0092-8674(00)80584-X Occurrence Handle1:CAS:528:DyaK1MXitFGhu7w%3D

    Article  PubMed  CAS  Google Scholar 

  • O Augusto MG Bonini AM Amanso E Linares CXC Santos S Lopes de Menezes (2002a) ArticleTitleNitrogen dioxide and carbonate radical anion: two emerging radicals in biology Free Rad Biol Med 32 841–859 Occurrence Handle10.1016/S0891-5849(02)00786-4 Occurrence Handle1:CAS:528:DC%2BD38XjtVeju7k%3D

    Article  CAS  Google Scholar 

  • O Augusto S Lopes de Menezes E Linares N Romero R Radi A Denicola (2002b) ArticleTitleEPR detection of glutathiyl and hemoglobin – cysteinyl radicals during the interaction of peroxynitrite with human erythrocytes Biochemistry 41 14323–14328 Occurrence Handle10.1021/bi0262202 Occurrence Handle1:CAS:528:DC%2BD38XosFSnsrY%3D

    Article  CAS  Google Scholar 

  • O Augusto MG Bonini DF Trindade (2004) ArticleTitleSpin trapping of glutathiyl and protein radicals produced from nitric oxide-derived oxidants Free Rad Biol Med 36 1224–1232 Occurrence Handle15110387 Occurrence Handle10.1016/j.freeradbiomed.2004.02.076 Occurrence Handle1:CAS:528:DC%2BD2cXjsVelsL0%3D

    Article  PubMed  CAS  Google Scholar 

  • C Badouard M Masuda H Nishino J Cadet A Favier JL Ravanat (2005) ArticleTitleDetection of chlorinated DNA and RNA nucleosides by HPLC coupled to tandem mass spectrometry as potential biomarkers of inflammation JChromatogr B Analyt Technol Biomed Life Sci 827 26–31 Occurrence Handle10.1016/j.jchromb.2005.03.025 Occurrence Handle1:CAS:528:DC%2BD2MXhtFOmsbjK

    Article  CAS  Google Scholar 

  • TP Barrows B Bhaskar TL Poulos (2004) ArticleTitleElectrostatic control of the tryptophan radical in cytochrome c peroxidase Biochemistry 43 8826–8834 Occurrence Handle15236591 Occurrence Handle10.1021/bi049531g Occurrence Handle1:CAS:528:DC%2BD2cXksl2kuro%3D

    Article  PubMed  CAS  Google Scholar 

  • MG Bonini O Augusto (2001) ArticleTitleCarbon dioxide stimulates the production of thiyl, sulfinyl, and disulfide radical anion from thiol oxidation by peroxynitrite J Biol Chem 276 9749–9754 Occurrence Handle11134018 Occurrence Handle10.1074/jbc.M008456200 Occurrence Handle1:CAS:528:DC%2BD3MXis1OlurY%3D

    Article  PubMed  CAS  Google Scholar 

  • MG Bonini S Miyamoto P Di Mascio O Augusto (2004a) ArticleTitleProduction of the carbonate radical anion during xanthine oxidase turnover in the presence of bicarbonate J Biol Chem 279 51836–51843 Occurrence Handle10.1074/jbc.M406929200 Occurrence Handle1:CAS:528:DC%2BD2cXhtVCktbvN

    Article  CAS  Google Scholar 

  • MG Bonini DC Fernandes O Augusto (2004b) ArticleTitleAlbumin oxidation to diverse radicals by the peroxidase activity of Cu,Zn-superoxide dismutase in the presence of bicarbonate or nitrite: diffusible radicals produce cysteinyl and solvent-exposed and -unexposed tyrosyl radicals Biochemistry 43 344–351 Occurrence Handle10.1021/bi035606p Occurrence Handle1:CAS:528:DC%2BD3sXpvVSmsrY%3D

    Article  CAS  Google Scholar 

  • NS Bryan T Rassaf RE Maloney CM Rodriguez F Saijo JR Rodriguez M Feelisch (2004) ArticleTitleCellular targets and mechanisms of nitros(yl)ation: An insight into their nature and kinetics in vivo Proc Natl Acad Sci USA 101 4308–4313 Occurrence Handle15014175 Occurrence Handle10.1073/pnas.0306706101 Occurrence Handle1:CAS:528:DC%2BD2cXivFartbo%3D

    Article  PubMed  CAS  Google Scholar 

  • S Carballal R Radi MC Kirk S Barnes BA Freeman B Alvarez (2003) ArticleTitleSulfenic acid formation in human serum albumin by hydrogen peroxide and peroxynitrite Biochemistry 42 9906–9914 Occurrence Handle12924939 Occurrence Handle10.1021/bi027434m Occurrence Handle1:CAS:528:DC%2BD3sXlvVyks7w%3D

    Article  PubMed  CAS  Google Scholar 

  • AL Clairbone H Miller D Parsonage PR Ross (1993) ArticleTitleProtein-sulfenic acid stabilization and function in enzyme catalysis and gene regulation FASEB J 7 1483–1490

    Google Scholar 

  • I Dalle-Donne A Scaloni D Giustarini E Cavarra G Tell G Lungarella R Colombo R Rossi A Milzani (2004) ArticleTitleProteins as biomarkers of oxidative/nitrosative stress in diseases: The contribution of redox proteomics Mass Spectrom Rev 24 55–99 Occurrence Handle10.1002/mas.20006

    Article  Google Scholar 

  • MJ Davies CL Hawkins (2004) ArticleTitleEPR spin-trapping of protein radicals Free Radic Biol Med 36 1072–1086 Occurrence Handle15082061 Occurrence Handle10.1016/j.freeradbiomed.2003.12.013 Occurrence Handle1:CAS:528:DC%2BD2cXjtVemt74%3D

    Article  PubMed  CAS  Google Scholar 

  • MJ Davies S Fu H Wang RT Dean (1999) ArticleTitleStable markers of oxidant damage to proteins and their application in the study of human disease Free Radic Biol Med 27 1151–1163 Occurrence Handle10641706 Occurrence Handle10.1016/S0891-5849(99)00206-3 Occurrence Handle1:CAS:528:DC%2BD3cXot1Gm

    Article  PubMed  CAS  Google Scholar 

  • LJ Deterding DC Ramirez JR Dubin RP Mason KB Tomer (2004) ArticleTitleIdentification of free radicals on hemoglobin from its self-peroxidation using mass spectrometry and immuno-spin trapping: observation of a histidinyl radical J Biol Chem 279 11600–11607 Occurrence Handle14699100 Occurrence Handle10.1074/jbc.M310704200 Occurrence Handle1:CAS:528:DC%2BD2cXitFyhtb4%3D

    Article  PubMed  CAS  Google Scholar 

  • JS Elam K Malek JA Rodriguez PA Doucette AB Taylor LJ Hayward DE Cabelli JS Valentine PJ Hart (2003) ArticleTitleAn alternative mechanism of bicarbonate-mediated peroxidation by copper-zinc superoxide dismutase: rates enhanced via proposed enzyme-associated peroxycarbonate intermediate J Biol Chem 278 21032–21039 Occurrence Handle12649272 Occurrence Handle10.1074/jbc.M300484200 Occurrence Handle1:CAS:528:DC%2BD3sXkt1Witrg%3D

    Article  PubMed  CAS  Google Scholar 

  • MG Espey DD Thomas KM Miranda DA Wink (2002) ArticleTitleFocusing of nitric oxide mediated nitrosation and oxidative nitrosylation as a consequence of reaction with superoxide Proc Natl Acad Sci USA 99 11127–11132 Occurrence Handle12177414 Occurrence Handle10.1073/pnas.152157599 Occurrence Handle1:CAS:528:DC%2BD38XmslSmtrc%3D

    Article  PubMed  CAS  Google Scholar 

  • DC Fernandes DB Medinas MJM Alves O Augusto (2005) ArticleTitleTempol diverts peroxynitrite/carbon dioxide reactivity toward albumin and cells from protein-tyrosine nitration to protein-cysteine nitrosation Free Radic Biol Med 38 189–200 Occurrence Handle15607902 Occurrence Handle10.1016/j.freeradbiomed.2004.09.027 Occurrence Handle1:CAS:528:DC%2BD2cXhtVyqsb7P

    Article  PubMed  CAS  Google Scholar 

  • T Finkel NJ Holbrook (2000) ArticleTitleOxidants, oxidative stress and the biology of ageing Nature 408 239–247 Occurrence Handle11089981 Occurrence Handle10.1038/35041687 Occurrence Handle1:CAS:528:DC%2BD3cXotFGltb0%3D

    Article  PubMed  CAS  Google Scholar 

  • RM Gatti R Radi O Augusto (1994) ArticleTitlePeroxynitrite-mediated oxidation of albumin to the protein thiyl free radical FEBS Lett 348 287–290 Occurrence Handle8034056 Occurrence Handle10.1016/0014-5793(94)00625-3 Occurrence Handle1:CAS:528:DyaK2cXlslegtrg%3D

    Article  PubMed  CAS  Google Scholar 

  • G Georgiou L Masip (2003) ArticleTitleAn overoxidation journey with a return ticket Science 300 592–594 Occurrence Handle12714731 Occurrence Handle10.1126/science.1084976 Occurrence Handle1:CAS:528:DC%2BD3sXktl2hsLc%3D

    Article  PubMed  CAS  Google Scholar 

  • DL Gilbert CA Colton (1999) Reactive oxygen species in biological systems: an interdisciplinary approach Kluwer Academic New York

    Google Scholar 

  • B Halliwell JMC Gutteridge (Eds) (1999) Free radicals in biology and medicine Oxford University Press New York

    Google Scholar 

  • J Heo G Gao SL Campbell (2004) ArticleTitlepH-dependent perturbation of Ras-guanine nucleotide interactions and Ras guanine nucleotide exchange Biochemistry 43 10102–10111 Occurrence Handle15287738 Occurrence Handle10.1021/bi035704a Occurrence Handle1:CAS:528:DC%2BD2cXlslers74%3D

    Article  PubMed  CAS  Google Scholar 

  • J Heo KC Prutzman V Mocanu SL Campbell (2005a) ArticleTitleMechanism of free radical nitric oxide-mediated Ras guanine nucleotide dissociation J Mol Biol 346 1423–1440 Occurrence Handle10.1016/j.jmb.2004.12.050 Occurrence Handle1:CAS:528:DC%2BD2MXhtlOksb8%3D

    Article  CAS  Google Scholar 

  • J Heo R Thapar SL Campbell (2005b) ArticleTitleRecognition and activation of Rho GTPases by Vav1 and Vav2 guanine nucleotide exchange factors Biochemistry 44 6573–6585 Occurrence Handle10.1021/bi047443q Occurrence Handle1:CAS:528:DC%2BD2MXjtVWgu7Y%3D

    Article  CAS  Google Scholar 

  • LJ Ignarro (1990) ArticleTitleBiosynthesis and metabolism of endothelium-derived nitric oxide Annu Rev Pharmacol Toxicol 30 535–560 Occurrence Handle2188578 Occurrence Handle10.1146/annurev.pa.30.040190.002535 Occurrence Handle1:CAS:528:DyaK3MXitFakug%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • D Jourd’heuil FL Jourd’heuil M Feelisch (2003) ArticleTitleOxidation and nitrosation of thiols at low micromolar exposure to nitric oxide. Evidence for a free radical mechanism J Biol Chem 278 15720–15726 Occurrence Handle12595536 Occurrence Handle10.1074/jbc.M300203200 Occurrence Handle1:CAS:528:DC%2BD3sXjt1eiu7Y%3D

    Article  PubMed  CAS  Google Scholar 

  • MB Kadiiska BC Gladen DD Baird D Germolec LB Graham CE Parker A Nyska JT Wachsman BN Ames S Basu N Brot GA Fitzgerald RA Floyd M George JW Heinecke GE Hatch K Hensley JA Lawson LJ Marnett JD Morrow DM Murray J Plastaras LJ Roberts Suffix2nd J Rokach MK Shigenaga RS Sohal J Sun RR Tice DH Van Thiel D Wellner PB Walter KB Tomer RP Mason JC Barrett (2005a) ArticleTitleBiomarkers of oxidative stress study II: are oxidation products of lipids, proteins, and DNA markers of CCl4 poisoning? Free Radic Biol Med 38 698–710 Occurrence Handle10.1016/j.freeradbiomed.2004.09.017 Occurrence Handle1:CAS:528:DC%2BD2MXhsVyqsLo%3D

    Article  CAS  Google Scholar 

  • MB Kadiiska BC Gladen DD Baird LB Graham CE Parker BN Ames S Basu GA Fitzgerald JA Lawson LJ Marnett JD Morrow DM Murray J Plastaras LJ Roberts Suffix2nd J Rokach MK Shigenaga J Sun PB Walter KB Tomer JC Barrett RP Mason (2005b) ArticleTitleBiomarkers of oxidative stress study III. Effects of the nonsteroidal anti-inflammatory agents indomethacin and meclofenamic acid on measurements of oxidative products of lipids in CCl4 poisoning Free Radic Biol Med 38 711–718 Occurrence Handle10.1016/j.freeradbiomed.2004.10.024 Occurrence Handle1:CAS:528:DC%2BD2MXhsVyqsLY%3D

    Article  CAS  Google Scholar 

  • KT Knecht RP Mason (1993) ArticleTitleIn vivo spin-trapping of xenobiotic free radical metabolites Arch Biochem Biophys 303 185–194 Occurrence Handle8390216 Occurrence Handle10.1006/abbi.1993.1272 Occurrence Handle1:CAS:528:DyaK3sXltlSkt78%3D

    Article  PubMed  CAS  Google Scholar 

  • PM Kris-Etherton AH Lichtenstein BV Howard D Steinberg JL Witztum InstitutionalAuthorNameNutrition Committee of the American Heart Association Council onNutrition, Physical Activity, and Metabolism (2004) ArticleTitleAntioxidant vitamin supplements and cardiovascular disease Circulation 110 637–641 Occurrence Handle15289389 Occurrence Handle10.1161/01.CIR.0000137822.39831.F1 Occurrence Handle1:CAS:528:DC%2BD2cXlvF2htbk%3D

    Article  PubMed  CAS  Google Scholar 

  • SI Liochev I Fridovich (2005) ArticleTitleThe role of CO2 in cobalt-catalyzed peroxidations Arch Biochem Biophys 439 99–104 Occurrence Handle15946639 Occurrence Handle10.1016/j.abb.2005.05.011 Occurrence Handle1:CAS:528:DC%2BD2MXlt1Gkur0%3D

    Article  PubMed  CAS  Google Scholar 

  • SI Liochev I Fridovich (2004) ArticleTitleCarbon dioxide mediates Mn(II)-catalyzed decomposition of hydrogen peroxide and peroxidation reactions Proc Natl Acad Sci USA 101 12485–12490 Occurrence Handle15310847 Occurrence Handle10.1073/pnas.0404911101 Occurrence Handle1:CAS:528:DC%2BD2cXnsVens7c%3D

    Article  PubMed  CAS  Google Scholar 

  • RP Mason (2004) ArticleTitleUsing anti-5,5-dimethyl-1-pyrroline N-oxide (anti-DMPO) to detect protein radicals in time and space with immuno-spin trapping Free Radic Biol Med 36 1214–1223 Occurrence Handle15110386 Occurrence Handle10.1016/j.freeradbiomed.2004.02.077 Occurrence Handle1:CAS:528:DC%2BD2cXjsVelsLw%3D

    Article  PubMed  CAS  Google Scholar 

  • S Moncada RM Palmer EA Higgs (1991) ArticleTitleNitric oxide: physiology, pathophysiology, and pharmacology Pharmacol Rev 43 109–142 Occurrence Handle1852778 Occurrence Handle1:CAS:528:DyaK3MXmt1amtLw%3D

    PubMed  CAS  Google Scholar 

  • JD Morrow (2005) ArticleTitleQuantification of isoprostanes as indices of oxidant stress and the risk of atherosclerosis in humans Arterioscler Thromb Vasc Biol 25 279–286 Occurrence Handle15591226 Occurrence Handle10.1161/01.ATV.0000152605.64964.c0 Occurrence Handle1:CAS:528:DC%2BD2MXmtFClug%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • R Radi A Denicola B Alvarez G Ferrer-Sueta H Rubbo (2000) The biological chemistry of peroxynitrite LJ Ignarro (Eds) Nitric oxide: biology and pathobiology Academic Press New York 57–89

    Google Scholar 

  • R Radi (2004) ArticleTitleNitric oxide, oxidants, and protein tyrosine nitration Proc Natl Acad Sci USA 101 4003–4008 Occurrence Handle15020765 Occurrence Handle10.1073/pnas.0307446101 Occurrence Handle1:CAS:528:DC%2BD2cXivFarsL8%3D

    Article  PubMed  CAS  Google Scholar 

  • DC Ramirez SE Mejiba RP Mason (2005) ArticleTitleCopper-catalyzed protein oxidation and its modulation by carbon dioxide: enhancement of protein radicals in cells J Biol Chem 280 27402–27411 Occurrence Handle15905164 Occurrence Handle10.1074/jbc.M504241200 Occurrence Handle1:CAS:528:DC%2BD2MXmt1Ggu74%3D

    Article  PubMed  CAS  Google Scholar 

  • DE Richardson CA Regino H Yao JV Johnson (2003) ArticleTitleMethionine oxidation by peroxymonocarbonate, a reactive oxygen species formed from CO2/bicarbonate and hydrogen peroxide Free Radic Biol Med 35 1538–1550 Occurrence Handle14680677 Occurrence Handle10.1016/j.freeradbiomed.2003.08.019 Occurrence Handle1:CAS:528:DC%2BD3sXpvVenurg%3D

    Article  PubMed  CAS  Google Scholar 

  • DE Richardson H Yao KM Frank DA Bennett (2000) ArticleTitleEquilibria, kinetics, and mechanisms in the bicarbonate activation of hydrogen peroxide: oxidation of sulfites by peroxymonocarbonate J Am Chem Soc 122 1729–1739 Occurrence Handle10.1021/ja9927467 Occurrence Handle1:CAS:528:DC%2BD3cXpsV2mtw%3D%3D

    Article  CAS  Google Scholar 

  • N Romero R Radi E Linares O Augusto CP Detweiler RP Mason A Denicola (2003) ArticleTitleReaction of human hemoglobin with peroxynitrite: isomerization to nitrate and secondary formation of protein radicals J Biol Chem 278 44049–44057 Occurrence Handle12920120 Occurrence Handle10.1074/jbc.M305895200 Occurrence Handle1:CAS:528:DC%2BD3sXoslWmsbg%3D

    Article  PubMed  CAS  Google Scholar 

  • H Sies R Mehlhom (1986) ArticleTitleMutagenicity of nitroxide-free radicals Arch Biochem Biophys 251 393–396 Occurrence Handle3539021 Occurrence Handle10.1016/0003-9861(86)90087-1 Occurrence Handle1:CAS:528:DyaL2sXhsVCksQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • MH Shishehbor RJA Aviles ML Brennan X Fu M Goormastic GL Pearce N Gokce JF Keaney MS Penn DL Sprecher JA Vita SL Hazen (2003) ArticleTitleAssociation of nitrotyrosine levels with cardiovascular disease and modulation by statin therapy J Am Med Assoc 289 1675–1680 Occurrence Handle10.1001/jama.289.13.1675 Occurrence Handle1:CAS:528:DC%2BD3sXisF2is7s%3D

    Article  CAS  Google Scholar 

  • JS Stamler (1994) ArticleTitleRedox signaling: nitrosylation and related target interactions of nitric oxide Cell 78 931–936 Occurrence Handle7923362 Occurrence Handle10.1016/0092-8674(94)90269-0 Occurrence Handle1:STN:280:ByqD3cjns1A%3D

    Article  PubMed  CAS  Google Scholar 

  • JS Stamler (1995) ArticleTitleS-Nitrosothiols and the bioregulatory actions of nitrogen oxides through reactions with thiol groups Curr Top Microbiol Immunol 196 19–36 Occurrence Handle7634823 Occurrence Handle1:CAS:528:DyaK2MXotl2rt74%3D

    PubMed  CAS  Google Scholar 

  • JR Stone (2004) ArticleTitleAn assessment of proposed mechanisms for sensing hydrogen peroxide in mammalian systems Arch Biochem Biophys 422 119–124 Occurrence Handle14759598 Occurrence Handle10.1016/j.abb.2003.12.029 Occurrence Handle1:CAS:528:DC%2BD2cXpsVKksw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • J Stubbe W van der Donk (1998) ArticleTitleProtein radicals in enzyme catalysis Chem Rev 98 705–762 Occurrence Handle11848913 Occurrence Handle10.1021/cr9400875 Occurrence Handle1:CAS:528:DyaK1cXhs1GrtbY%3D

    Article  PubMed  CAS  Google Scholar 

  • M Torres HJ Forman (2003) ArticleTitleRedox signaling and the MAP kinase pathways Biofactors 17 287–296 Occurrence Handle12897450 Occurrence Handle1:CAS:528:DC%2BD3sXlsl2rt7c%3D

    PubMed  CAS  Google Scholar 

  • J Vasquez-Vivar AM Santos VBC Junqueira O Augusto (1996) ArticleTitlePeroxynitrite-mediated formation of free radicals in human plasma: EPR detection of ascorbyl, albumin-thiyl and uric acid-derived free radicals Biochem J 314 869–876 Occurrence Handle8615782 Occurrence Handle1:CAS:528:DyaK28XhvFyjt78%3D

    PubMed  CAS  Google Scholar 

  • HA Woo HZ Chae SC Hwang K-S Yang SW Kang K Kim SG Rhee (2003) ArticleTitleReversing the inactivation of peroxiredoxins caused by cysteine sulfinic acid formation Science 300 653–656 Occurrence Handle12714748 Occurrence Handle10.1126/science.1080273 Occurrence Handle1:CAS:528:DC%2BD3sXjtVylsr0%3D

    Article  PubMed  CAS  Google Scholar 

  • ZA Wood LB Poole PA Karplus (2003) ArticleTitlePeroxiredoxin evolution and the regulation of hydrogen peroxide signaling Science 300 650–653 Occurrence Handle12714747 Occurrence Handle10.1126/science.1080405 Occurrence Handle1:CAS:528:DC%2BD3sXjtVylsr8%3D

    Article  PubMed  CAS  Google Scholar 

  • H Yao DE Richardson (2003) ArticleTitleBicarbonate surfoxidants: micellar oxidations of aryl sulfides with bicarbonate-activated hydrogen peroxide J Am Chem Soc 125 6211–6221 Occurrence Handle12785853 Occurrence Handle10.1021/ja0274756 Occurrence Handle1:CAS:528:DC%2BD3sXjt1Shtb4%3D

    Article  PubMed  CAS  Google Scholar 

  • H Zhang C Andrekopoulos J Joseph J Crow B Kalyanaraman (2004) ArticleTitleThe carbonate radical anion-induced covalent aggregation of human copper, zinc superoxide dismutase, and alpha-synuclein: intermediacy of tryptophan- and tyrosine-derived oxidation Free Radic Biol Med 36 1355–1365 Occurrence Handle15135171 Occurrence Handle10.1016/j.freeradbiomed.2004.02.038 Occurrence Handle1:CAS:528:DC%2BD2cXjvVagsr8%3D

    Article  PubMed  CAS  Google Scholar 

  • H Zhang Y Xu J Joseph B Kalyanaraman (2005) ArticleTitleIntramolecular electron transfer between tyrosyl radical and cysteine residue inhibits tyrosine nitration and induces thiyl radical formation in model peptides treated with myeloperoxidase, H2O2, and NO2 −: EPR spin trapping studies J Biol Chem 280 40684–40698 Occurrence Handle16176930 Occurrence Handle10.1074/jbc.M504503200 Occurrence Handle1:CAS:528:DC%2BD2MXht1OqtrfK

    Article  PubMed  CAS  Google Scholar 

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  1. Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil

    O. Augusto & S. Muntz Vaz

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Augusto, O., Muntz Vaz, S. EPR spin-trapping of protein radicals to investigate biological oxidative mechanisms. Amino Acids 32, 535–542 (2007). https://doi.org/10.1007/s00726-006-0429-4

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