Research Article
No access
Published Online: 8 January 2010

Edaravone Inhibits Protein Carbonylation by a Direct Carbonyl-Scavenging Mechanism: Focus on Reactivity, Selectivity, and Reaction Mechanisms

Publication: Antioxidants & Redox Signaling
Volume 12, Issue Number 3

Abstract

The aim of this study was to evaluate the ability of the well-known radical scavenging compound edaravone (EDA) to entrap and detoxify reactive carbonyl species (RCS) derived from lipid peroxidation [4-hydroxy-trans-2-nonenal (HNE), acrolein and glyoxal], as well as its ability to prevent RCS-induced protein carbonylation, by using hemoglobin (Hb) modified by HNE as an in vitro model. Through a combined HPLC and high-resolution mass spectrometric approach, we confirmed the ability of EDA to scavenge precursors for either advanced glycation or lipoxidation end products (EAGLEs), such as glyoxal, and demonstrated for the first time that EDA is also a potent quencher of α,β-unsaturated aldehydes (providing mass spectral characterization of the adducts), being significantly more active than a series of well-known RCS sequestering agents. Direct infusion analysis of the intact protein and nano LC-ESI-MS/MS analysis of the tryptic digest, carried out on an LTQ-Orbitrap hybrid mass spectrometer, were used to study the modifications occurring on Hb after exposure to increasing HNE concentrations, providing evidence for Cys93 (Hb β-chain) involvement in covalent attachment, and to demonstrate the ability of EDA dose-dependently to inhibit Hb carbonylation. Computational studies allowed us to elucidate the mechanism of EDA-RCS interaction and to explain the preferential site of HNE adduction to Hb. The same combined approach indicated that EDA is not a selective RCS scavenger, being able to react also with nontoxic, physiologically relevant aldehydes, such as pyridoxal. Antioxid. Redox Signal. 12, 381–392.

Get full access to this article

View all available purchase options and get full access to this article.

References

1.
Aldini GDalle-Donne IColombo RMaffei Facino RMilzani ACarini M. Lipoxidation-derived reactive carbonyl species as potential drug targets in preventing protein carbonylation and related cellular dysfunctionChem Med Chem11045-10582006. 1. Aldini G, Dalle-Donne I, Colombo R, Maffei Facino R, Milzani A, and Carini M. Lipoxidation-derived reactive carbonyl species as potential drug targets in preventing protein carbonylation and related cellular dysfunction. Chem Med Chem 1: 1045–1058, 2006.
2.
Aldini GDalle-Donne IFacino RMMilzani ACarini M. Intervention strategies to inhibit protein carbonylation by lipoxidation-derived reactive carbonylsMed Res Rev27817-1682007. 2. Aldini G, Dalle-Donne I, Facino RM, Milzani A, and Carini M. Intervention strategies to inhibit protein carbonylation by lipoxidation-derived reactive carbonyls. Med Res Rev 27: 817–168, 2007.
3.
Balawender RKomorowski L. Atomic Fukui function indices and local softness ab initioJ Chem Phys1095203-52111998. 3. Balawender R and Komorowski L. Atomic Fukui function indices and local softness ab initio. J Chem Phys 109: 5203–5211, 1998.
4.
Butterfield DACastegna A. Proteomics for the identification of specifically oxidized proteins in brain: technology and application to the study of neurodegenerative disordersAmino Acids25419-4252003. 4. Butterfield DA and Castegna A. Proteomics for the identification of specifically oxidized proteins in brain: technology and application to the study of neurodegenerative disorders Amino Acids 25: 419–425, 2003.
5.
Butterfield DASultana RPoon HFDalle-Donne IScaloni AButterfield DA. Redox proteomics: a new approach to investigate oxidative stress in Alzheimer's diseaseRedox Proteomics: From Protein Modifications to Cellular Dysfunction and DiseaseHobokenWiley2006563-603. 5. Butterfield DA, Sultana R, and Poon HF. Redox proteomics: a new approach to investigate oxidative stress in Alzheimer's disease. In: Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Disease, edited by Dalle-Donne I, Scaloni A, and Butterfield DA. Hoboken: Wiley, 2006, pp. 563–603.
6.
Carini MAldini GMaffei Facino R. Mass spectrometry for detection of 4-hydroxy-trans-2-nonenal (HNE) adducts with peptides and proteinsMass Spectrom Rev23281-3052004. 6. Carini M, Aldini G, and Maffei Facino R. Mass spectrometry for detection of 4-hydroxy-trans-2-nonenal (HNE) adducts with peptides and proteins. Mass Spectrom Rev 23: 281–305, 2004.
7.
Crabb JWO'Neil JMiyagi MWest KHoff HF. Hydroxynonenal inactivates cathepsin B by forming Michael adducts with active site residuesProtein Sci11831-8402002. 7. Crabb JW, O'Neil J, Miyagi M, West K, and Hoff HF. Hydroxynonenal inactivates cathepsin B by forming Michael adducts with active site residues. Protein Sci 11: 831–840, 2002.
8.
Dalle-Donne IRossi RGiustarini DMilzani AColombo R. Protein carbonyl groups as biomarkers of oxidative stressClin Chim Acta32923-382003. 8. Dalle-Donne I, Rossi R, Giustarini D, Milzani A, and Colombo R. Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta 329: 23–38, 2003.
9.
Dalle-Donne IScaloni AGiustarini DCavarra ETell GLungarella GColombo RRossi RMilzani A. Proteins as biomarkers of oxidative/nitrosative stress in diseases: the contribution of redox proteomicsMass Spectrom Rev2455-992005. 9. Dalle-Donne I, Scaloni A, Giustarini D, Cavarra E, Tell G, Lungarella G, Colombo R, Rossi R, and Milzani A. Proteins as biomarkers of oxidative/nitrosative stress in diseases: the contribution of redox proteomics. Mass Spectrom Rev 24: 55–99, 2005.
10.
Dalle-Donne IAldini GCarini MColombo RRossi RMilzani A. Protein carbonylation, cellular dysfunction, and disease progressionJ Cell Mol Med10389-4062006. 10. Dalle-Donne I, Aldini G, Carini M, Colombo R, Rossi R, and Milzani A. Protein carbonylation, cellular dysfunction, and disease progression. J Cell Mol Med 10: 389–406, 2006.
11.
Domingo LRBenchoukb WMekellecheb SM. Understanding the role of the Lewis acid catalyst on the 1,3-dipolar cycloaddition of N-benzylideneaniline N-oxide with acrolein: a DFT studyTetrahedron634464-44712007. 11. Domingo LR, Benchoukb W, and Mekellecheb SM. Understanding the role of the Lewis acid catalyst on the 1,3-dipolar cycloaddition of N-benzylideneaniline N-oxide with acrolein: a DFT study. Tetrahedron 63: 4464–4471, 2007.
12.
Fenaille FGuy PATabet JC. Study of protein modification by 4-hydroxy-2-nonenal and other short-chain aldehydes analyzed by electrospray ionization tandem mass spectrometryJ Am Soc Mass Spectrom14215-2262003. 12. Fenaille F, Guy PA, and Tabet JC. Study of protein modification by 4-hydroxy-2-nonenal and other short-chain aldehydes analyzed by electrospray ionization tandem mass spectrometry. J Am Soc Mass Spectrom 14: 215–226, 2003.
13.
Higashi Y. Edaravone for the treatment of acute cerebral infarction: role of endothelium-derived nitric oxide and oxidative stressExpert Opin Pharmacother10323-3312009. 13. Higashi Y. Edaravone for the treatment of acute cerebral infarction: role of endothelium-derived nitric oxide and oxidative stress. Expert Opin Pharmacother 10: 323–331, 2009.
14.
Izuhara YNangaku MTakizawa STakahashi SShao JOishi HKobayashi Hvan Ypersele de Strihou CMiyata T. A novel class of advanced glycation inhibitors ameliorates renal and cardiovascular damage in experimental rat modelsNephrol Dial Transplant23497-5092008. 14. Izuhara Y, Nangaku M, Takizawa S, Takahashi S, Shao J, Oishi H, Kobayashi H, van Ypersele de Strihou C, and Miyata T. A novel class of advanced glycation inhibitors ameliorates renal and cardiovascular damage in experimental rat models. Nephrol Dial Transplant 23: 497–509, 2008.
15.
LoPachin RMBarber DSGavin T. Synaptosomal toxicity and nucleophilic targets of 4-hydroxy-2-nonenalToxicol Sci104235-2492008. 15. LoPachin RM, Barber DS, and Gavin T. Synaptosomal toxicity and nucleophilic targets of 4-hydroxy-2-nonenal. Toxicol Sci 104: 235–249, 2008.
16.
Miyata Tvan Ypersele de Strihou CUeda YIchimori KInagi ROnogi HIshikawa NNangaku MKurokawa K. Angiotensin II receptor antagonists and angiotensin-converting enzyme inhibitors lower in vitro the formation of advanced glycation end products: biochemical mechanismsJ Am Soc Nephrol132478-24872002. 16. Miyata T, van Ypersele de Strihou C, Ueda Y, Ichimori K, Inagi R, Onogi H, Ishikawa N, Nangaku M, and Kurokawa K. Angiotensin II receptor antagonists and angiotensin-converting enzyme inhibitors lower in vitro the formation of advanced glycation end products: biochemical mechanisms. J Am Soc Nephrol 13: 2478–2487, 2002.
392
17.
Negre-Salvayre ACoatrieux CIngueneau CSalvayre R. Advanced lipid peroxidation end products in oxidative damage to proteins: potential role in diseases and therapeutic prospects for the inhibitorsBr J Pharmacol1536-202008. 17. Negre-Salvayre A, Coatrieux C, Ingueneau C, and Salvayre R. Advanced lipid peroxidation end products in oxidative damage to proteins: potential role in diseases and therapeutic prospects for the inhibitors. Br J Pharmacol 153: 6–20, 2008.
18.
Nielsen JE. Analysing the pH-dependent properties of proteins using pKa calculationsJ Mol Graph Model25691-6992007. 18. Nielsen JE. Analysing the pH-dependent properties of proteins using pKa calculations. J Mol Graph Model 25: 691–699, 2007.
19.
Nyström T. Role of oxidative carbonylation in protein quality control and senescenceEMBO J241311-13172005. 19. Nyström T. Role of oxidative carbonylation in protein quality control and senescence. EMBO J 24: 1311–1317, 2005.
20.
O'Brien PJSiraki AGShangari N. Aldehyde sources, metabolism, molecular toxicity mechanisms, and possible effects on human healthCrit Rev Toxicol35609-6622005. 20. O'Brien PJ, Siraki AG, and Shangari N. Aldehyde sources, metabolism, molecular toxicity mechanisms, and possible effects on human health. Crit Rev Toxicol 35: 609–662, 2005.
21.
Olsen JVde Godoy LMLi GMacek BMortensen PPesch RMakarov ALange OHorning SMann M. Parts per million mass accuracy on an Orbitrap mass spectrometer via lock mass injection into a C-trapMol Cell Proteomics122010-20212005. 21. Olsen JV, de Godoy LM, Li G, Macek B, Mortensen P, Pesch R, Makarov A, Lange O, Horning S, and Mann M. Parts per million mass accuracy on an Orbitrap mass spectrometer via lock mass injection into a C-trap. Mol Cell Proteomics 12: 2010–2021, 2005.
22.
Pedretti AVilla LVistoli G. Modeling of binding modes and inhibition mechanism of some natural ligands of farnesyl transferase using molecular dockingJ Med Chem451460-14652002. 22. Pedretti A, Villa L, and Vistoli G. Modeling of binding modes and inhibition mechanism of some natural ligands of farnesyl transferase using molecular docking. J Med Chem 45: 1460–1465, 2002.
23.
Pennathur SHeinecke JW. Mechanisms for oxidative stress in diabetic cardiovascular diseaseAntioxid Redox Signal9955-9692007. 23. Pennathur S and Heinecke JW. Mechanisms for oxidative stress in diabetic cardiovascular disease. Antioxid Redox Signal 9: 955–969, 2007.
24.
Poli GSchaur RJSiems WGLeonarduzzi G. 4-Hydroxynonenal: a membrane lipid oxidation product of medicinal interestMed Res Rev28569-6312008. 24. Poli G, Schaur RJ, Siems WG, and Leonarduzzi G. 4-Hydroxynonenal: a membrane lipid oxidation product of medicinal interest. Med Res Rev 28: 569–631, 2008.
25.
Rees MSvan Kuijk FJGMSiakotos ANMundy BP. Improved synthesis of various isotope labelled 4-hydroxyalkenals and peroxidation intermediatesSynth Commun253225-32281995. 25. Rees MS, van Kuijk FJGM, Siakotos AN, and Mundy BP. Improved synthesis of various isotope labelled 4-hydroxyalkenals and peroxidation intermediates. Synth Commun 25: 3225–3228, 1995.
26.
Kind TFiehn O. Seven golden rules for heuristic filtering of molecular formulas obtained by accurate mass spectrometryBMC Bioinformatics8105-1212007. 26. Kind T and Fiehn O. Seven golden rules for heuristic filtering of molecular formulas obtained by accurate mass spectrometry. BMC Bioinformatics 8: 105–121, 2007.
27.
Uchida K. Role of reactive aldehyde in cardiovascular diseasesFree Radic Biol Med281685-16962000. 27. Uchida K. Role of reactive aldehyde in cardiovascular diseases. Free Radic Biol Med 28: 1685–1696, 2000.
28.
Vistoli GOrioli MPedretti ARegazzoni LCanevotti RNegrisoli GCarini MAldini G. Design, synthesis, and evaluation of carnosine derivatives as selective and efficient sequestering agents of cytotoxic reactive carbonyl speciesChem Med Chem4967-9752009. 28. Vistoli G, Orioli M, Pedretti A, Regazzoni L, Canevotti R, Negrisoli G, Carini M, and Aldini G. Design, synthesis, and evaluation of carnosine derivatives as selective and efficient sequestering agents of cytotoxic reactive carbonyl species. Chem Med Chem 4: 967–975, 2009
29.
Watanabe TTahara MTodo S. The novel antioxidant edaravone: from bench to bedsideCardiovasc Ther26101-1142008. 29. Watanabe T, Tahara M, and Todo S. The novel antioxidant edaravone: from bench to bedside. Cardiovasc Ther 26: 101–114, 2008.
30.
Xi HAkishita MNagai KYu WHasegawa HEto MKozaki KToba K. Potent free radical scavenger, edaravone, suppresses oxidative stress-induced endothelial damage and early atherosclerosisAtherosclerosis191281-2892007. 30. Xi H, Akishita M, Nagai K, Yu W, Hasegawa H, Eto M, Kozaki K, and Toba K. Potent free radical scavenger, edaravone, suppresses oxidative stress-induced endothelial damage and early atherosclerosis. Atherosclerosis 191: 281–289, 2007.
31.
Yocum AKYergey ALBlair IA. Novel lipid hydroperoxide-derived hemoglobin histidine adducts as biomarkers of oxidative stressJ Mass Spectrom40754-7642005. 31. Yocum AK, Yergey AL, and Blair IA. Novel lipid hydroperoxide-derived hemoglobin histidine adducts as biomarkers of oxidative stress. J Mass Spectrom 40: 754–764, 2005.
32.
Zhang NKomine-Kobayashi MTanaka RLiu MMizuno YUrabe T. Edaravone reduces early accumulation of oxidative products and sequential inflammatory responses after transient focal ischemia in mice brainStroke32220-22252005. 32. Zhang N, Komine-Kobayashi M, Tanaka R, Liu M, Mizuno Y, and Urabe T. Edaravone reduces early accumulation of oxidative products and sequential inflammatory responses after transient focal ischemia in mice brain. Stroke 3: 2220–2225, 2005.

Information & Authors

Information

Published In

cover image Antioxidants & Redox Signaling
Antioxidants & Redox Signaling
Volume 12Issue Number 3February 1, 2010
Pages: 381 - 392
PubMed: 19722825

History

Published in print: February 1, 2010
Published online: 8 January 2010
Published ahead of print: 30 October 2009
Published ahead of production: 1 September 2009
Accepted: 15 August 2009
Received: 7 August 2009

Permissions

Request permissions for this article.

Authors

Affiliations

Giancarlo Aldini
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi,” Università degli Studi di Milano, Milan, Italy.
Giulio Vistoli
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi,” Università degli Studi di Milano, Milan, Italy.
Luca Regazzoni
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi,” Università degli Studi di Milano, Milan, Italy.
Maria Carmela Benfatto
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi,” Università degli Studi di Milano, Milan, Italy.
Ilaria Bettinelli
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi,” Università degli Studi di Milano, Milan, Italy.
Marina Carini
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi,” Università degli Studi di Milano, Milan, Italy.

Notes

Address correspondence to:
Marina Carini
Dipartimento di Scienze Farmaceutiche “Pietro Pratesi”
Università degli Studi di Milano
Via L. Mangiagalli 25
20133 Milan,
Italy
E-mail: [email protected]

Author Disclosure Statement

No competing financial interests exist.

Metrics & Citations

Metrics

Citations

Export citation

Select the format you want to export the citations of this publication.

View Options

Get Access

Access content

To read the fulltext, please use one of the options below to sign in or purchase access.

Society Access

If you are a member of a society that has access to this content please log in via your society website and then return to this publication.

Restore your content access

Enter your email address to restore your content access:

Note: This functionality works only for purchases done as a guest. If you already have an account, log in to access the content to which you are entitled.

View options

PDF/EPUB

View PDF/ePub

Full Text

View Full Text

Media

Figures

Other

Tables

Share

Share

Copy the content Link

Share on social media

Back to Top