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The Dynamics of the Formation of Cytochrome c Complexes with Anionic Lipids and the Mechanism of the Production of Lipid Radicals Catalyzed by These Complexes

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Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology Aims and scope

Abstract

In the present work, the recorded kinetics of chemiluminescence enhanced by coumarin 334 indicated that the complexes of cytochrome c (Cyt) with anionic lipids (Cyt-AL), such as cardiolipin (CL) and phosphatidic acid (PA), when added to mitochondrial and cytoplasmic membranes, are capable of catalysing reactions of lipid free radical formation. The effect of Cyt-CL complex was studied on mitochondria isolated from the liver of male Wistar rats and male outbred mice, as well as on erythrocytes isolated from the blood of male Wistar rats. It was shown that complex Cyt-PA exerts a peroxidase effect similar to the effect produced by Cyt-CL. Upon exposure to Cyt-CL, the formation of free radicals was more prominent in mitochondrial ghosts (mitochondria subjected to freeze-thawing and washed by several precipitation–resuspension cycles) than in intact mitochondria. This suggests that Cyt-CL does not pass through biological membranes: even if it penetrates into them, it probably gets stuck in the phospholipid bilayer. Thus, the previously discovered cytotoxic effect of Cyt-CL on cancer cells is most likely due to peroxidation of the cytoplasmic membrane, but not of the inner membrane of mitochondria.

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ACKNOWLEDGMENTS

The work was supported by the Russian Science Foundation (project no. 17-74-10248).

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Authors and Affiliations

  1. Shubnikov Institute of Crystallography of Federal Scientific Research Center Crystallography and Photonics, Russian Academy of Sciences, 119333, Moscow, Russia

    G. K. Vladimirov, A. M. Nesterova, A. A. Levkina, M. V. Kovalchuk & Yu. A. Vladimirov

  2. Institute for Regenerative Medicine, Sechenov University, 119991, Moscow, Russia

    G. K. Vladimirov, A. M. Nesterova & Yu. A. Vladimirov

  3. Pirogov Russian National Research Medical University, 117997, Moscow, Russia

    A. A. Levkina, A. N. Osipov & Y. O. Teselkin

  4. National Research Center Kurchatov Institute, 123182, Moscow, Russia

    M. V. Kovalchuk

  5. Moscow Lomonosov State University, 119991, Moscow, Russia

    Yu. A. Vladimirov

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  1. G. K. Vladimirov
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  2. A. M. Nesterova
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  3. A. A. Levkina
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Correspondence to G. K. Vladimirov.

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The authors declare that they have no conflict of interests.

All experiments with animals were conducted in accordance with the regulations established by Order of the USSR Ministry of Health of 08/12/1977, no. 755, as well as European Communities Council Directive 1986 (86/609/EEC).

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Translated by G. Vladimirov

Abbreviations: AL, anionic lipids; C-334, coumarin 334; ChL, chemiluminescence; CytC, cytochrome c; Cyt-AL, cytochrome c complex with anionic lipids; CL, cardiolipin; Cyt-CL, complex cytochrome c with cardiolipin; Cyt-TOCL, a complex of cytochrome c with tetraoleoyl cardiolipin; LOO·, lipoperoxyl radical; LOOH, lipid hydroperoxides; PA, dioleoyl-phosphatidic acid; TOCL, tetraoleoyl cardiolipin.

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Vladimirov, G.K., Nesterova, A.M., Levkina, A.A. et al. The Dynamics of the Formation of Cytochrome c Complexes with Anionic Lipids and the Mechanism of the Production of Lipid Radicals Catalyzed by These Complexes. Biochem. Moscow Suppl. Ser. A 14, 232–241 (2020). https://doi.org/10.1134/S1990747820030137

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