Biochemistry (Moscow)

, Volume 74, Issue 4, pp 438–444 | Cite as

Electrochemistry of cytochromes P450: Analysis of current-voltage characteristics of electrodes with immobilized cytochromes P450 for the screening of substrates and inhibitors

  • V. V. ShumyantsevaEmail author
  • T. V. Bulko
  • G. P. Kuznetsova
  • N. F. Samenkova
  • A. I. Archakov


In the current study, an approach to elucidating the substrate specificity of cytochromes P450 based on the analysis of current-voltage characteristics of voltammograms and amperograms is proposed. Data on the electrochemical behavior of bioelectrodes with immobilized cytochromes P450 2B4, 1A2, 3A4, 11A1 (P450scc), and 51b1 (Mycobacterium tuberculosis sterol 14α-demethylase or CYP51 MT) in the presence of typical substrates and inhibitors for these hemoprotein forms are reported. Immobilization of the enzymes was accomplished by using graphite screen-printed electrodes modified with gold nanoparticles and with the synthetic membrane-like compound didodecyldimethylammonium bromide. The method of electro-analysis can be applied to the search of potential substrates and inhibitors of cytochromes P450 and to creation of multichannel electrochemical plates (chips, panels) with immobilized cytochromes P450.

Key words

cytochrome P450 colloidal gold nanoparticles bioelectrochemistry drug metabolism 





cyclic voltammetry


didodecyldimethylammonium bromide


differential pulse voltammetry

Epa and Epc

potentials of anode reduction and cathode oxidation peaks, respectively


intermittent pulse amperometry


square wave voltammetry




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Omura, T., and Sato, R. (1962) J. Biol. Chem., 237, 1375–1376.PubMedGoogle Scholar
  2. 2.
    Nebert, D. W., and Russell, D. W. (2002) The Lancet, 360, 1155–1162.CrossRefGoogle Scholar
  3. 3.
    Kumar, R. A., and Clark, D. S. (2006) Curr. Opin. Chem. Biol., 10, 162–168.PubMedCrossRefGoogle Scholar
  4. 4.
    Chen, H., and Morgan, J. A. (2206) Enzyme Microb. Technol., 38, 760–764.CrossRefGoogle Scholar
  5. 5.
    Lee, M.-Y., Park, C. B., Dordick, J. S., and Clark, D. S. (2005) Proc. Natl. Acad. Sci. USA, 102, 983–987.PubMedCrossRefGoogle Scholar
  6. 6.
    Sakai-Kato, K., Kato, M., Homma, H., Toyo’oka, T., and Utsunomiya-Tate, N. (2005) Anal. Chem., 77, 7080–7083.PubMedCrossRefGoogle Scholar
  7. 7.
    Turpeinen, M., Korhonen, L. E., Tolonen, A., Uusitalo, J., Juvonen, R., Raunio, H., and Felkonen, O. (2006) Eur. J. Pharm Sci., 29, 130–138.PubMedCrossRefGoogle Scholar
  8. 8.
    Estabrook, R. W., Faulkner, K. M., Shet, M. S., and Fisher, C. W. (1996) Meth. Enzymol., 272(B), 44–51.PubMedCrossRefGoogle Scholar
  9. 9.
    Bistolas, N., Wollenberger, U., Jung, C., and Scheller, F. W. (2005) Biosens. Bioelectron., 20, 2408–2423.PubMedCrossRefGoogle Scholar
  10. 10.
    Shumyantseva, V. V., Bulko, T. V., and Archakov, A. I. (2005) J. Inorg. Biochem., 99, 1051–1063.PubMedCrossRefGoogle Scholar
  11. 11.
    Udit, A. K., and Gray, H. B. (2005) Biochem. Biophys. Res. Commun., 338, 470–476.PubMedCrossRefGoogle Scholar
  12. 12.
    Bakker, E., and Telting-Diaz, M. (2002) Anal. Chem., 74, 2781–2800.PubMedCrossRefGoogle Scholar
  13. 13.
    Mehrvar, M., and Abdi, M. (2004) Anal. Sci., 20, 1113–1126.PubMedCrossRefGoogle Scholar
  14. 14.
    Wang, J. (2005) Analyst, 130, 421–426.PubMedCrossRefGoogle Scholar
  15. 15.
    Murphy, L. (2006) Curr. Opin. Chem. Biol., 10, 177–184.PubMedCrossRefGoogle Scholar
  16. 16.
    Pumera, M., Sanchez, S., Ichnose, I., and Tang, J. (2007) Sensors and Actuators, 123, 1195–1205.CrossRefGoogle Scholar
  17. 17.
    Tansil, N., and Gao, Z. (2006) Nanotoday, 1, 28–37.Google Scholar
  18. 18.
    Shumyantseva, V. V., Bulko, T. V., Rudakov, Yu. O., Kuznetsova, G. P., Samenkova, N. F., Lisitsa, A. V., Karuzina, I. I., and Archakov, A. I. (2007) J. Inorg. Biochem., 101, 859–865.PubMedCrossRefGoogle Scholar
  19. 19.
    Berthell, D., Brust, M., Schiffrin, D. J., and Kiely, C. (1996) J. Electroanal. Chem., 409, 137–143.CrossRefGoogle Scholar
  20. 20.
    Han, X., Cheng, W., Zhang, Z., Dong, S., and Wang, E. (2002) Biochim. Biophys. Acta, 1556, 273–277.PubMedCrossRefGoogle Scholar
  21. 21.
    Karuzina, I. I., Zgoda, V. G., Kuznetsova, G. P., Samenkova, N. F., and Archakov, A. I. (1999) Free Rad. Biol. Med., 26, 620–632.PubMedCrossRefGoogle Scholar
  22. 22.
    Omura, T., and Sato, R. (1964) J. Biol. Chem., 239, 2379–2385.PubMedGoogle Scholar
  23. 23.
    Imai, Y., Hashimoto, C., Satake, H., Girardin, A., and Sato, R. (1980) J. Biochem., 88, 489–503.PubMedGoogle Scholar
  24. 24.
    Hashimoto, C., and Imai, Y. (1976) Biochem. Biophys. Res. Commun., 68, 821–827.PubMedCrossRefGoogle Scholar
  25. 25.
    Shavkunov, A. S., Lazarev, V. N., Zgoda, V. G., Govorun, V. M., Levi, P., Yanssen, P., and Archakov, A. I. (2003) Biomed. Khim., 49, 145–152.PubMedGoogle Scholar
  26. 26.
    Nash, T. (1953) Biochem. J., 55, 416–421.PubMedGoogle Scholar
  27. 27.
    Kanaeva, I. P., Dedinskii, I. R., Scotselyas, E. D., Krainev, A. G., Guleva, I. V., Sevryukova, I. F., Koen, Ya. M., Kuznetsova, G. P., Bachmanova, G. I., and Archakov, A. I. (1992) Arch. Biochem. Biophys., 298, 395–412.PubMedCrossRefGoogle Scholar
  28. 28.
    Shumyantseva, V. V., Bulko, T. V., Rudakov, Yu. O., Kuznetsova, G. P., Samenkova, N. F., Lisitsa, A. V., Karuzina, I. I., and Archakov, A. I. (2007) Biochemistry (Moscow), Suppl. Ser. B: Biomed. Chem., 1, 327–333.CrossRefGoogle Scholar
  29. 29.
    Shumyantseva, V. V., Bulko, T. V., Kuznetsova, G. P., Lisitsa, A. V., Ponomarenko, E. A., Karuzina, I. I., and Archakov, A. I. (2007) Biochemistry (Moscow), 72, 658–663.CrossRefGoogle Scholar
  30. 30.
    Smith, D., Sadagopan, N., Zientek, M., Reddy, A., and Cohen, L. (2007) J. Chromatogr. B, 850, 455–463.CrossRefGoogle Scholar
  31. 31.
    Persson, K. P., Ekehed, S., Otter, C., Lutz, M., McPheat, J., Masimirembwa, C. M., and Andersson, T. B. (2006) Pharm. Res., 23, 56–69.PubMedCrossRefGoogle Scholar
  32. 32.
    Tupeinen, M., Jouko, U., Jorma, J., and Olavi, P. (2005) Eur. J. Pharm. Res., 24, 123–132.CrossRefGoogle Scholar
  33. 33.
    Johnson, D. L., Lewis, B. C., Elliot, D. J., Miners, J. O., and Martin, L. L. (2005) Biochem. Pharmacol., 69, 1533–1541.PubMedCrossRefGoogle Scholar
  34. 34.
    Shukla, A., Gillam, E. M., Mitchll, D. J., and Bernhardt, P. V. (2005) Electrochem. Commun., 7, 437–442.CrossRefGoogle Scholar
  35. 35.
    Shumyantseva, V. V., Ivanov, Yu. D., Bistolas, N., Scheller, F. W., Archakov, A. I., and Wollenberger, U. (2004) Anal. Chem., 76, 6046–6052.PubMedCrossRefGoogle Scholar
  36. 36.
    Lei, C., Wollenberger, U., Jung, C., and Scheller, F. W. (2000) Biochem. Biophys. Res. Commun., 268, 740–744.PubMedCrossRefGoogle Scholar
  37. 37.
    Shumyantseva, V. V., Deluca, J., Bulko, T. V., Carrara, S., Nicolini, C., Usanov, S. A., and Archakov, A. I. (2004) Biosens. Bioelectron., 19, 971–976.PubMedCrossRefGoogle Scholar
  38. 38.
    Shumyantseva, V. V., Bavastrello, V., Carrara, S., Riley, D. J., Bulko, T. V., Skryabin, K. G., Archakov, A. I., and Nicolini, C. (2005) Biosens. Bioelectron., 21, 217–222.PubMedCrossRefGoogle Scholar
  39. 39.
    Joseph, S., Rusling, J. F., Lvov, Y. M., Friedberg, T., and Fuhr, U. (2003) Biochem. Pharmacol., 65, 1817–1826.PubMedGoogle Scholar
  40. 40.
    Lepesheva, G. I., Strushkevich, N. V., and Usanov, S. A. (1999) Biochim. Biophys. Acta, 1434, 31–43.PubMedGoogle Scholar
  41. 41.
    Lepesheva, G. I., and Waterman, M. R. (2004) Mol. Cell. Endocrinol., 215, 165–170.PubMedCrossRefGoogle Scholar
  42. 42.
    Ortiz de Montellano, P. R. (ed.) (1995) Cytochrome P450, 2nd Edn., Plenum Press, New York, pp. 307–317.Google Scholar
  43. 43.
    Lewis, D. F. V. (2001) Guide to Cytochrome P450. Structure and Function (Taylor and Francis, eds.) London-New York, pp. 90–93.Google Scholar
  44. 44.
    Kitson, T. M. (1996) Bioorg. Chem., 24, 331–339.CrossRefGoogle Scholar
  45. 45.
    Prough, R. A., Burke, M. D., and Mayger, R. T. (1978) Meth. Enzymol., 52, 372–377.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • V. V. Shumyantseva
    • 1
    Email author
  • T. V. Bulko
    • 1
  • G. P. Kuznetsova
    • 1
  • N. F. Samenkova
    • 1
  • A. I. Archakov
    • 1
  1. 1.Institute of Biomedical ChemistryRussian Academy of Medical SciencesMoscowRussia

Personalised recommendations