Skip to main content
SpringerLink
Log in

Detection of Peroxynitrite-Induced Protein and DNA Modifications

  • Protocol
Book cover Oxidants and Antioxidants

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 196))

Abstract

Nitric oxide and superoxide rapidly combine to form a toxic reaction product, peroxynitrite anion (ONOO) (1,2). The oxidant reactivity of peroxynitrite is mediated by an intermediate with the biological activity of the hydroxyl radical. However, this product does not appear to be the hydroxyl radical per se, but peroxynitrous acid (ONOOH) or its activated isomer (ONOOH*) (2). Peroxynitrite readily reacts with proteins, lipids, and DNA under conditions of inflammation.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Beckman, J. S., Beckman, T. W., Chen, J., Marshall, P. A., and Freeman, B. A. (1990) Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc. Natl. Acad. Sci. USA 87, 1620–1624.

    Article  PubMed  CAS  Google Scholar 

  2. Pryor, W. A. and Squadrito, G. L. (1995) The chemistry of peroxynitrite: a product from the reaction of nitric oxide with superoxide. Am. J. Physiol. 268, L699–L722.

    PubMed  CAS  Google Scholar 

  3. Ischiropoulos, H., Zhu, L., Chen, J., Tsai, M., Martin, J. C., Smith, C. D., and Beckman, J. S. (1992) Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase. Arch. Biochem. Biophys. 298, 431–437.

    Article  PubMed  CAS  Google Scholar 

  4. Szabo, C. and Ohshima, H. (1997) DNA damage induced by peroxynitrite: subsequent biological effects. Nitric Oxide 1, 373–385.

    Article  PubMed  CAS  Google Scholar 

  5. Szabó, C. and Dawson, V. L. (1998) Role of poly(ADP-ribose) synthetase in inflammation and ischaemia-reperfusion. Trends. Pharmacol. Sci. 19, 287–298.

    Article  PubMed  Google Scholar 

  6. Viera, L., Zu, Y., Eztevez, A. G., and Beckman J. S. (1999) Immunohistochemical methods to detect nitrotyrosine. Methods Enzymol. 301, 373–381.

    Article  PubMed  CAS  Google Scholar 

  7. Haddad, I. Y., Pataki, G., Hu, P., Galliani, C., Beckman, J. S., and Matalon S. (1994) Quantitation of nitrotyrosine levels in lung sections of patients and animals with acute lung injury. J. Clin. Invest. 94, 2407–2413.

    Article  PubMed  CAS  Google Scholar 

  8. Crowley, J. R., Yarasheski, K., Leeuwenburgh, C., Turk, J., and Heinecke, J. W. (1998) Isotope dilution mass spectrometric quantification of 3-nitrotyrosine in proteins and tissues is facilitated by reduction to 3-aminotyrosine. Anal. Biochem. 259, 127–135.

    Article  PubMed  CAS  Google Scholar 

  9. Souza, J. M., Daikhin, E., Yudkoff, M., Raman, C. S., and Ischiropoulos, H. (1999) Factors determining the selectivity of protein tyrosine nitration. Arch. Biochem. Biophys. 371, 169–178.

    Article  PubMed  CAS  Google Scholar 

  10. Banks, B. A., Ischiropoulos, H., McClelland, M., Ballard, P. L., and Ballard, R. A. (1998) Plasma 3-nitrotyrosine is elevated in premature infants who develop bronchoplumonary dysplasia. Pediatrics 101, 870–874.

    Article  PubMed  CAS  Google Scholar 

  11. Khan, J., Brennan, D. M., Bradley, N., Gao, B., Bruckdorfer, R., and Jacobs, M. (1997) 3-Nitrotyrosine in the proteins of human plasma determined by an ELISA method. Biochem. J. 330, 795–801.

    Google Scholar 

  12. Ter Steege, J. C. A., Koster-Kamphuis, L., van Straaten, E. A., Forget, P. P., and Buurman, W. A. (1998) Nitrotyrosine in plasma of celiac disease patients as detected by a new sandwich ELISA. Free Rad. Biol. Med. 25, 953–963.

    Article  PubMed  CAS  Google Scholar 

  13. MacMillan, L. A. and Thompson, J. A. (1999) Immunoprecipitation of nitro-tyrosine containing proteins. Methods Enzymol. 301, 135–145.

    Article  Google Scholar 

  14. Byun, J., Mueller, D. M., and Heinecke, J. W. (1999) 8-Nitro-2′-deoxyguanosine, a specific marker of oxidation by reactive nitrogen species, is generated by the myeloperoxidase-hydrogen peroxide-nitrite system of activated human phagocytes. Biochemistry 38, 2590–2600.

    Article  PubMed  CAS  Google Scholar 

  15. Ohshima, H., Celan, I., Chazotte, L., Pignatelli, B., and Mower, H. F. (1999) Analysis of 3-nitrotyrosine in biological fluids and protein hydrolyzates by high-performance liquid chromatography using a postseparation, on-line reduction column and electrochemical detection: results with various nitrating agents [In Process Citation]. Nitric Oxide 3, 132–141.

    Article  PubMed  CAS  Google Scholar 

  16. Shigenaga, M. K., Lee, H. H., Blount, B. C., Christen, S., Shigeno, E. T., Yip, H., and Ames, B. N. (1997) Inflammation and NO(X)-induced nitration: assay for 3-nitrotyrosine by HPLC with electrochemical detection. Proc. Natl. Acad. Sci. USA 94, 3211–3216.

    Article  PubMed  CAS  Google Scholar 

  17. Yermilov, V., Rubio, J., Becchi, M., Friesen, M. D., Pignatelli, B., and Ohshima, H. (1995) Formation of 8-nitroguanine by the reaction of guanine with peroxynitrite in vitro. Carcinogenesis 16, 2045–2050.

    Article  PubMed  CAS  Google Scholar 

  18. Byun, J., Mueller, D. M., and Heinecke, J. W. (1999) 8-Nitro-2′-deoxyguanosine, a specific marker of oxidation by reactive nitrogen species, is generated by the myeloperoxidase-hydrogen peroxide-nitrite system of activated human phagocytes. Biochemistry 38, 2590–2600.

    Article  PubMed  CAS  Google Scholar 

  19. Sodum, R. S., Nie, G., and Fiala, E. S. (1993) 8-Aminoguanine: a base modification produced in rat liver nucleic acids by the hepatocarcinogen 2-nitropropane. Chem. Res. Toxicol. 6, 269–276.

    Article  PubMed  CAS  Google Scholar 

  20. Yermilov, V., Rubio, J., and Ohshima, H. (1995) Formation of 8-nitroguanine in DNA treated with peroxynitrite in vitro and its rapid removal from DNA by depurination. FEBS Lett. 376, 207–210.

    Article  PubMed  CAS  Google Scholar 

  21. Reed, J. W., Ho, H. H., and Jolly, W. L. (1974) Chemical syntheses with a quenched flow reactor. Hydroxytrihydroborate and peroxynitrite. J. Am. Chem. Soc. 96, 1248–1249.

    Article  CAS  Google Scholar 

  22. de Murcia, G. and Menissier de Murcia, F. (1994) Poly(ADP-ribose) polymerase: a molecular nick-sensor. Trends. Biochem. Sci. 19, 172–176.

    Article  PubMed  Google Scholar 

  23. Rosenthal, D. S., Ding, R., Simbulan-Rosenthal, C. M., Vaillancourt, J. P., Nicholson, D. W., and Smulson, M. (1997) Intact cell evidence for the early synthesis, and subsequent late apopain-mediated suppression, of poly(ADP-ribose) during apoptosis. Exp. Cell Res. 232, 313–321.

    Article  PubMed  CAS  Google Scholar 

  24. Affar, E. B., Duriez, P. J., Shah, R. G., Winstall, E., Germain, M., Boucher, C., et al. (1999) Immunological determination and size characterization of poly(ADP-ribose) synthesized in vitro and in vivo. Biochim. Biophys. Acta 1428(2–3), 137–146.

    Google Scholar 

  25. Virág, L., Scott, G. S., Cuzzocrea, S., Marmer, D., Salzman, A. L., and Szabó, C. (1998) Peroxynitrite-induced thymocyte apoptosis: the role of caspases and poly (ADP-ribose) synthetase (PARS) activation. Immunology 94, 345–355.

    Article  PubMed  Google Scholar 

  26. Virág, L., Scott, G. S., Antal-Szalmás, P., O’Connor, M., Ohshima, H., and Szabó, C. (1999) Requirement of intracellular calcium mobilization for peroxynitrite-induced poly(ADP-ribose) synthetase activation and cytotoxicity. Mol. Pharmacol. 56, 824–833.

    PubMed  Google Scholar 

  27. Pulido, E. J., Shames, B. D., Selzman, C. H., Barton, H. A., Banerjee, A., Bensard, D. D., and McIntyre, R. C. (1999) Inhibition of PARS attenuates endotoxin-induced dysfunction of pulmonary vasorelaxation. Am. J. Physiol. 277 L769–L776.

    PubMed  CAS  Google Scholar 

  28. Kupper, J. H., Van Gool, L., Muller, M., and Burkle, A. (1996) Detection of poly(ADP-ribose) polymerase and its reaction product poly(ADP-ribose) by immunocytochemistry. Histochem. J. 28, 391–395.

    Article  PubMed  CAS  Google Scholar 

  29. Kupper, J. H., Muller, M., Jacobson, M. K., et al. (1995) Trans-dominant inhibi-tion of poly(ADP-ribosyl)ation sensitizes cells against gamma-irradiation and N-methyl-N’-nitro-N-nitrosoguanidine but does not limit DNA replication of a polyomavirus replicon. Mol. Cell Biol. 15, 3154–3163.

    PubMed  CAS  Google Scholar 

  30. Lankenau, S., Burkle, A., and Lankenau, D. H. (1999) Detection of poly (ADP-ribose) synthesis in Drosophila testes upon γ-irradiation. Chromosoma 108, 44–51.

    Article  PubMed  CAS  Google Scholar 

  31. Kawamitsu, H., Hoshino, H., Okada, H., Miwa, M., Momoi, H., and Sugimura, T. (1984) Monoclonal antibodies to poly(adenosine diphosphate ribose) recognize different structures. Biochemistry 23 (16), 3771–3777.

    Article  PubMed  CAS  Google Scholar 

  32. Love, S., Barber, R., and Wilcock, G. K. (1999) Increased poly(ADP-ribosyl)ation of nuclear proteins in Alzheimer’s disease. Brain 122, 247–253.

    Article  PubMed  Google Scholar 

  33. Scott, G. S., Jakeman, L. B., Stokes, B. T., and Szabó, C. (1999) Peroxynitrite production and activation of poly(ADP-ribose) synthetase in spinal cord injury. Ann. Neurol. 45, 120–124.

    Article  PubMed  CAS  Google Scholar 

  34. Garcia Soriano, F., Virag, L., Jagtap, P., et al. (2001) Diabetic endothelial dysfunc-tion: the role of poly(ADP-ribose) polymerase activation. Nat. Med. 7, 108–113.

    Article  PubMed  CAS  Google Scholar 

  35. Bakondi, E., Bai, P., Szabo, E., et al. (2002) Detection of poly(ADP-ribose) polymerase activation in oxidatively stressed cells and tissues using biotinylated NAD substrate. J. Histochem/Cytochem 50, 91–98.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Stokes Research Institute Children’s Hospital of Philadelphia, Department of Biochemistry and Biophysics, The University of Pennsylvania Medical Center, Philadelphia, PA

    Scott Lorch, Richard Lightfoot, Caryn Hertkorn, Marie Weiss & Harry Ischiropoulos

  2. Unit of Endogenous Cancer Risk Factors, International Agency for Research on Cancer, Lyon, France

    Hiroshi Ohshima, Jose Souza, Vladimir Yermilov, Brigitte Pignatelli & Mituharu Masuda

  3. Department of Medical Chemistry, Faculty of Medicine, University of Debrecen, Debrecen, Hungary

    László Virág

  4. Department of Biochemistry and Biophysics, Stokes Research Institute Children’s Hospital of Philadelphia, The University of Pennsylvania Medical Center, Philadelphia, PA

    Qiping Chen

  5. Inotek Corporation, Beverly, MA

    László Virág & Csaba Szabó

Authors
  1. Scott Lorch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Richard Lightfoot
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroshi Ohshima
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. László Virág
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qiping Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Caryn Hertkorn
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marie Weiss
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jose Souza
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Harry Ischiropoulos
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Vladimir Yermilov
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Brigitte Pignatelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Mituharu Masuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Csaba Szabó
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Florida, Gainesville, FL

    Donald Armstrong

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Humana Press Inc.

About this protocol

Cite this protocol

Lorch, S. et al. (2002). Detection of Peroxynitrite-Induced Protein and DNA Modifications. In: Armstrong, D. (eds) Oxidants and Antioxidants. Methods in Molecular Biology™, vol 196. Humana Press. https://doi.org/10.1385/1-59259-274-0:247

Download citation

  • DOI: https://doi.org/10.1385/1-59259-274-0:247

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-851-6

  • Online ISBN: 978-1-59259-274-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation