Biochemistry (Moscow)

, Volume 80, Issue 12, pp 1589–1597 | Cite as

Uncoupling and toxic action of alkyltriphenylphosphonium cations on mitochondria and the bacterium Bacillus subtilis as a function of alkyl chain length

  • L. S. Khailova
  • P. A. Nazarov
  • N. V. Sumbatyan
  • G. A. Korshunova
  • T. I. Rokitskaya
  • V. I. Dedukhova
  • Yu. N. AntonenkoEmail author
  • V. P. Skulachev


A series of permeating cations based on alkyl derivatives of triphenylphosphonium (Cn-TPP+) containing linear hydrocarbon chains (butyl, octyl, decyl, and dodecyl) was investigated in systems of isolated mitochondria, bacteria, and liposomes. In contrast to some derivatives (esters) of rhodamine-19, wherein butyl rhodamine possessed the maximum activity, in the case of Cn-TPP a stimulatory effect on mitochondrial respiration steadily increased with growing length of the alkyl radical. Tetraphenylphosphonium and butyl-TPP+ at a dose of several hundred micromoles exhibited an uncoupling effect, which might be related to interaction between Cn-TPP+ and endogenous fatty acids and induction of their own cyclic transfer, resulting in transport of protons across the mitochondrial membrane. Such a mechanism was investigated by measuring efflux of carboxyfluorescein from liposomes influenced by Cn-TPP+. Experiments with bacteria demonstrated that dodecyl-TPP+, decyl-TPP+, and octyl-TPP+ similarly to quinone-containing analog (SkQ1) inhibited growth of the Gram-positive bacterium Bacillus subtilis, wherein the inhibitory effect was upregulated with growing lipophilicity. These cations did not display toxic effect on growth of the Gram-negative bacterium Escherichia coli. It is assumed that the difference in toxic action on various bacterial species might be related to different permeability of bacterial coats for the examined triphenylphosphonium cations.


permeating cation SkQ1 mitochondria uncoupling bacteria cytotoxicity 





alkyl-triphenylphosphonium cation


potential-dependent carbocyanine probe


mitochondrial transmembrane proton electrochemical gradient


pH gradient across the inner membrane of mitochondria


mitochondrial membrane potential


arbonyl cyanide p-trifluoromethoxyphenylhydrazone


tetraphenylphosphonium cation




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  1. 1.
    Skulachev, V. P., Antonenko, Y. N., Cherepanov, D. A., Chernyak, B. V., Izyumov, D. S., Khailova, L. S., Klishin, S. S., Korshunova, G. A., Lyamzaev, K. G., Pletjushkina, O. Y., Roginsky, V. A., Rokitskaya, T. I., Severin, F. F., Severina, I. I., Simonyan, R. A., Skulachev, M. V., Sumbatyan, N. V., Sukhanova, E. I., Tashlitsky, V. N., Trendeleva, T. A., Vyssokikh, M. Y., and Zvyagilskaya, R. A. (2010. Prevention of cardiolipin oxidation and fatty acid cycling as two antioxidant mechanisms of cationic derivatives of plastoquinone (SkQs), Biochim. Biophys. Acta, 1797, 878–889.CrossRefPubMedGoogle Scholar
  2. 2.
    Murphy, M. P., and Smith, R. A. (2007. Targeting antioxidants to mitochondria by conjugation to lipophilic cations, Annu. Rev. Pharmacol. Toxicol., 47, 629–656.CrossRefPubMedGoogle Scholar
  3. 3.
    Cunha, F. M., Caldeira da Silva, C. C., Cerqueira, F. M., and Kowaltowski, A. J. (2011. Mild mitochondrial uncoupling as a therapeutic strategy, Curr. Drug Targets, 12, 783–789.CrossRefPubMedGoogle Scholar
  4. 4.
    Korshunov, S. S., Skulachev, V. P., and Starkov, A. A. (1997. High protonic potential actuates a mechanism of production of reactive oxygen species in mitochondria, FEBS Lett., 416, 15–18.CrossRefPubMedGoogle Scholar
  5. 5.
    Severin, F. F., Severina, I. I., Antonenko, Y. N., Rokitskaya, T. I., Cherepanov, D. A., Mokhova, E. N., Vyssokikh, M. Y., Pustovidko, A. V., Markova, O. V., Yaguzhinsky, L. S., Korshunova, G. A., Sumbatyan, N. V., Skulachev, M. V., and Skulachev, V. P. (2010. Penetrating cation/fatty acid anion pair as a mitochondria-targeted protonophore, Proc. Natl. Acad. Sci. USA, 107, 663–668.PubMedCentralCrossRefPubMedGoogle Scholar
  6. 6.
    Antonenko, Y. N., Avetisyan, A. V., Bakeeva, L. E., Chernyak, B. V., Chertkov, V. A., Domnina, L. V., Ivanova, O. Y., Izyumov, D. S., Khailova, L. S., Klishin, S. S., Korshunova, G. A., Lyamzaev, K. G., Muntyan, M. S., Nepryakhina, O. K., Pashkovskaya, A. A., Pletjushkina, O. Y., Pustovidko, A. V., Roginsky, V. A., Rokitskaya, T. I., Ruuge, E. K., Saprunova, V. B., Severina, I. I., Simonyan, R. A., Skulachev, I. V., Skulachev, M. V., Sumbatyan, N. V., Sviryaeva, I. V., Tashlitsky, V. N., Vassiliev, J. M., Vyssokikh, M. Y., Yaguzhinsky, L. S., Zamyatnin, A. A., Jr., and Skulachev, V. P. (2008. Mitochondria-targeted plastoquinone derivatives as tools to interrupt execution of the aging program. 1. Cationic plastoquinone derivatives: synthesis and in vitro studies, Biochemistry (Moscow), 73, 12731287.CrossRefGoogle Scholar
  7. 7.
    Khailova, L. S., Silachev, D. N., Rokitskaya, T. I., Avetisyan, A. V., Lyamsaev, K. G., Severina, I. I., Il’yasova, T. M., Gulyaev, M. V., Dedukhova, V. I., Trendeleva, T. A., Plotnikov, E. Y., Zvyagilskaya, R. A., Chernyak, B. V., Zorov, D. B., Antonenko, Y. N., and Skulachev, V. P. (2014. A short-chain alkyl derivative of rhodamine 19 acts as a mild uncoupler of mitochondria and a neuroprotector, Biochim. Biophys. Acta, 1837, 1739–1747.CrossRefPubMedGoogle Scholar
  8. 8.
    Sassi, N., Mattarei, A., Azzolini, M., Szabo’, I., Paradisi, C., Zoratti, M., and Biasutto, L. (2014. Cytotoxicity of mitochondria-targeted resveratrol derivatives: interactions with respiratory chain complexes and ATP synthase, Biochim. Biophys. Acta, 1837, 1781–1789.CrossRefPubMedGoogle Scholar
  9. 9.
    Severina, I. I., Muntyan, M. S., Lewis, K., and Skulachev, V. P. (2001. Transfer of cationic antibacterial agents berberine, palmatine, and benzalkonium through bimolecular planar phospholipid film and Staphylococcus aureus membrane, IUBMB Life, 52, 321–324.CrossRefPubMedGoogle Scholar
  10. 10.
    Schmeller, T., Latz-Bruning, B., and Wink, M. (1997. Biochemical activities of berberine, palmatine, and sanguinarine mediating chemical defence against microorganisms and herbivores, Phytochemistry, 44, 257–266.CrossRefPubMedGoogle Scholar
  11. 11.
    Galkina, I. V., and Egorova, S. N. (2009. Biological activity of quaternary salts of phosphonium and perspectives of their medical application, Farmatsiya, 9, 142–145.Google Scholar
  12. 12.
    Galkina, I. V., Bakhtiyarova, Y. V., Shulaeva, M. P., Pozdeev, O. K., Egorova, S. N., Cherkasov, R. A., and Galkin, V. I. (2013) Synthesis and antimicrobial activity of carboxylate phosphabetaines derivatives with alkyl chains of various lengths, J. Chem., doi: 10.1155/2013/302937.Google Scholar
  13. 13.
    Kanazawa, A., Ikeda, T., and Endo, T. (1994. Synthesis and antimicrobial activity of dimethyl-substituted and trimethylsubstituted phosphonium salts with alkyl chains of various lengths, Antimicrob. Agents Chemother., 38, 945–952.PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Ross, M. F., Prime, T. A., Abakumova, I., James, A. M., Porteous, C. M., Smith, R. A., and Murphy, M. P. (2008. Rapid and extensive uptake and activation of hydrophobic triphenylphosphonium cations within cells, Biochem. J., 411, 633–645.CrossRefPubMedGoogle Scholar
  15. 15.
    Rokitskaya, T. I., Sumbatyan, N. V., Tashlitsky, V. N., Korshunova, G. A., Antonenko, Y. N., and Skulachev, V. P. (2010. Mitochondria-targeted penetrating cations as carriers of hydrophobic anions through lipid membranes, Biochim. Biophys. Acta, 1798, 1698–1706.CrossRefPubMedGoogle Scholar
  16. 16.
    Johnson, D., and Lardy, H. (1967. Isolation of liver or kidney mitochondria, Methods Enzymol., 10, 94–96.CrossRefGoogle Scholar
  17. 17.
    Akerman, K. E., and Wikstrom, M. K. (1976. Safranine as a probe of the mitochondrial membrane potential, FEBS Lett., 68, 191–197.CrossRefPubMedGoogle Scholar
  18. 18.
    Miller, J. B., and Koshland, D. E., Jr. (1977. Sensory electrophysiology of bacteria: relationship of the membrane potential to motility and chemotaxis in Bacillus subtilis, Proc. Natl. Acad. Sci. USA, 74, 4752–4756.PubMedCentralCrossRefPubMedGoogle Scholar
  19. 19.
    Trendeleva, T. A., Rogov, A. G., Cherepanov, D. A., Sukhanova, E. I., Il’yasova, T. M., Severina, I. I., and Zvyagilskaya, R. A. (2012. Interaction of tetraphenylphosphonium and dodecyltriphenylphosphonium with lipid membranes and mitochondria, Biochemistry (Moscow), 77, 1021–1028.CrossRefGoogle Scholar
  20. 20.
    Lotscher, H. R., Winterhalter, K. H., Carafoli, E., and Richter, C. (1980. The energy state of mitochondria during the transport of Ca2+, Eur. J. Biochem., 110, 211–216.CrossRefPubMedGoogle Scholar
  21. 21.
    Kramer, R., and Palmieri, F. (1989. Molecular aspects of isolated and reconstituted carrier proteins from animal mitochondria, Biochim. Biophys. Acta, 974, 1–23.CrossRefPubMedGoogle Scholar
  22. 22.
    Yang, Q., Liu, X. Y., Umetani, K., Kamo, N., and Miyake, J. (1999. Partitioning of triphenylalkylphosphonium homologues in gel bead-immobilized liposomes: chromatographic measurement of their membrane partition coefficients, Biochim. Biophys. Acta, 1417, 122–130.CrossRefPubMedGoogle Scholar
  23. 23.
    Bakeeva, L. E., Grinius, L. L., Jasaitis, A. A., Kuliene, V. V., Levitsky, D. O., Liberman, E. A., Severina, I. I., and Skulachev, V. P. (1970. Conversion of biomembrane-produced energy into electric form. II. Intact mitochondria, Biochim. Biophys Acta, 216, 13–21.CrossRefPubMedGoogle Scholar
  24. 24.
    Thorsteinsson, T., Loftsson, T., and Masson, M. (2003. Soft antibacterial agents, Curr. Med. Chem., 10, 1129–1136.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  • L. S. Khailova
    • 1
  • P. A. Nazarov
    • 1
  • N. V. Sumbatyan
    • 2
  • G. A. Korshunova
    • 1
  • T. I. Rokitskaya
    • 1
  • V. I. Dedukhova
    • 1
  • Yu. N. Antonenko
    • 1
    Email author
  • V. P. Skulachev
    • 1
    • 3
  1. 1.Lomonosov Moscow State UniversityBelozersky Institute of Physico-Chemical BiologyMoscowRussia
  2. 2.Lomonosov Moscow State UniversityFaculty of ChemistryMoscowRussia
  3. 3.Lomonosov Moscow State UniversityInstitute of MitoengineeringMoscowRussia

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