Applied Magnetic Resonance

, 28:411 | Cite as

Detection of free radicals formation in haemolymph of insects by EPR spectroscopy

  • D. A. Komarov
  • I. A. Slepneva
  • V. V. Glupov
  • V. V. Khramtsov


The generation of reactive oxygen species during oxidation of DOPA (3-(3,4-dihydroxyphenyl-DL-alanine) in haemolymph of insects has been studied by electron paramagnetic resonance with spin traps 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide and 1-hydroxy-3-carboxy-pyrrolidine. The data support the formation of DOPA-derived highly reactive intermediates during melanization in haemolymph of insects. The formation of superoxide has not been detected apparently due to reaction of DOPA with superoxide. The rate constant of the latter reaction was estimated ask≈5·105 M−1.c−1. The formation of DOPA-semiquinone in haemolymph has been measured using spin stabilization ofo-semiquinones by Mg2+. The comparative studies in haemolymph of intact insects and insects infected by fungal infection suggest an important role of DOPA-semiquinone production in the immune status of insects.


Electron Paramagnetic Resonance Electron Paramagnetic Resonance Spectrum Dopa Electron Paramagnetic Resonance Signal Electron Paramagnetic Resonance Spectroscopy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Ashida M., Brey P.T. in: Molecular Mechanisms of Immune Responses in Insects (Brey P.T., Hultmark D., eds.), pp. 135–172. London: Chapman and Hall 1997.Google Scholar
  2. 2.
    Johansson M.W., Söderhäll K. in: Invertebrate Immunology (Rinkevich B., Müller W.E.G., eds.), pp. 46–66. Berlin. Springer 1995.Google Scholar
  3. 3.
    Slepneva I.A., Glupov V.V., Sergeeva S.V., Khramtsov V.V.: Biochem. Biophys. Res. Commun.264, 212–215 (1999)CrossRefGoogle Scholar
  4. 4.
    Slepneva I.A., Komarov D.A., Glupov V.V., Serebrov V.V., Khramtsov V.V.: Biochem. Biophys. Res. Commun.300, 188–191 (2003)CrossRefGoogle Scholar
  5. 5.
    Nappi A.J., Vass E.: Pigment Cell Res.6, 17–126 (1993)CrossRefGoogle Scholar
  6. 6.
    Nappi A.J., Ottavianni E.: Bio Essays22, 469–480 (2000)Google Scholar
  7. 7.
    Arakawa T.: J. Insect Physiol.40, 165–171 (1994)CrossRefGoogle Scholar
  8. 8.
    Nappi A.J., Vass E., Frey F., Carton Y.: Eur. J. Cell Biol.68, 450–456 (1995)Google Scholar
  9. 9.
    Nappi A.J., Vass E.: J. Parasitol.84, 1150–1157 (1998)CrossRefGoogle Scholar
  10. 10.
    Glupov V.V., Khvoshevskaya M.F., Lozinskaya Ya.L., Dubovski I.M., Martemyanov V.V., Sokolova Ju.Ya.: Cytobios106, 165–178 (2001)Google Scholar
  11. 11.
    Goettel M.S., Inglis G.D. in: Manual of Techniques in Insect Pathology (Lacey L., ed.), pp. 213–279. New York: Academic Press 1997.CrossRefGoogle Scholar
  12. 12.
    Kim C., DiCosimo R., Filippo J.S.: Anal. Chem.51, 679–681 (1979)CrossRefGoogle Scholar
  13. 13.
    Frejaville C., Karoui H., Tuccio B., Le Moigne F., Culcasi M., Pietri S., Lauricella R., Tordo P.: J. Med. Chem.38, 258–265 (1995)CrossRefGoogle Scholar
  14. 14.
    Dikalov S., Skatchkov M., Bassenge E.: Biochem. Biophys. Res. Commun.231, 701–704 (1997)CrossRefGoogle Scholar
  15. 15.
    Eaton D.R.: Inorg. Chem.3, 1268–1271 (1964)CrossRefGoogle Scholar
  16. 16.
    Kalyanaraman B., Sealy R.C.: Biochem. Biophys. Res. Commun.106, 1119–1125 (1982)CrossRefGoogle Scholar
  17. 17.
    Bielski B.H.J., Cabelli D.E., Arudi R.L., Ross A.B.: J. Phys. Chem. Ref. Data14, 1041–1100 (1985)ADSGoogle Scholar
  18. 18.
    Dikalov S., Skatchkov M., Fink B., Bassenge E.: Nitric Oxide1, 423–431 (1997)CrossRefGoogle Scholar
  19. 19.
    Felix C.C., Sealy R.C.: J. Am. Chem. Soc.103, 2831–2836 (1981)CrossRefGoogle Scholar
  20. 20.
    Kalyanaraman B., Felix C.C., Sealy R.C.: J. Biol. Chem.259, 7584–7589 (1984)Google Scholar
  21. 21.
    Kalyanaraman B.: Methods Enzymol.186, 333–343 (1990)CrossRefGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • D. A. Komarov
    • 1
  • I. A. Slepneva
    • 1
  • V. V. Glupov
    • 2
  • V. V. Khramtsov
    • 1
    • 3
  1. 1.Institute of Chemical Kinetics and CombustionRussian Academy of SciencesNovosibirskRussian Federation
  2. 2.Institute of Animal Systematics and EcologyRussian Academy of SciencesNovosibirskRussian Federation
  3. 3.Dorothy M. Davis Heart and Lung Research InstituteThe Ohio State UniversityColumbusUSA

Personalised recommendations