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A Comparative Study of the Action of Protonophore Uncouplers and Decoupling Agents as Inducers of Free Respiration in Mitochondria in States 3 and 4: Theoretical and Experimental Approaches

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Abstract

Theoretical and experimental studies have revealed that that in the liver mitochondria an increase in the rate of free respiration in state 3 induced by protonophore uncouplers 2,4-dinitrophenol and сarbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone is equal to or slightly greater than the increase in respiration rate in state 4 induced by these uncouplers. In contrast to these protonophore uncouplers, the decoupler α,ω-tetradecanedioic acid, increasing the rate of respiration in state 4, does not significantly affect the rate of free respiration in state 3. We have proposed quantitative indicators that allow determining the constituent part of the rate of respiration in state 4, associated with the decoupling effect of the uncoupler. Using the example of palmitic acid, we have found out the fundamental possibility of the simultaneous functioning of uncouplers by two mechanisms: as protonophores and as decouplers. The data obtained contradict the delocalized version of Mitchell’s chemiosmotic theory, but are in complete agreement with its local version. It can be assumed that the F0F1-ATP synthase and nearby respiratory chain complexes form a local zone of coupled respiration and oxidative ATP synthesis (zones of oxidative phosphorylation). The uncoupler-induced stimulation of mitochondrial free respiration of mitochondria in state 3 is mainly due to the return of protons to the matrix in local zones, where the generation of a proton motive force (Δр) by respiratory chain complexes is associated with various transport processes, but not with ATP synthesis (zones of protonophore uncoupling). In contrast, respiratory stimulation in state 4 by decouplers is realized in local zones of oxidative phosphorylation by switching the respiratory chain complexes to the idle mode of operation in the absence of ATP synthesis.

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Abbreviations

DNP:

2,4-dinitrophenol

FCCP:

сarbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone

TDA:

α,ω-tetradecanedioic acid

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Acknowledgements

The work was supported by grant from the Russian Foundation for Basic Research (20-015-00124) and by the grant of the President of the Russian Federation (MK-61.2019.4).

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  1. Mari State University, pl. Lenina 1, Mari El, Yoshkar-Ola, 424001, Russia

    Victor N. Samartsev, Alena A. Semenova & Mikhail V. Dubinin

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  1. Victor N. Samartsev
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  2. Alena A. Semenova
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  3. Mikhail V. Dubinin
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Correspondence to Mikhail V. Dubinin.

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Statement on the welfare of animals. All procedures were performed in accordance with the European Communities Council Directive (November 24, 1986; 86/609/EEC) and the Declaration on humane treatment of animals. The Protocol of experiments was approved by the Mari State University Ethics Committee.

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Samartsev, V.N., Semenova, A.A. & Dubinin, M.V. A Comparative Study of the Action of Protonophore Uncouplers and Decoupling Agents as Inducers of Free Respiration in Mitochondria in States 3 and 4: Theoretical and Experimental Approaches. Cell Biochem Biophys 78, 203–216 (2020). https://doi.org/10.1007/s12013-020-00914-5

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