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Interaction of Venturicidin and Fo·F1-ATPase/ATP Synthase of Tightly Coupled Subbacterial Particles of Paracoccus denitrificans in Energized Membranes

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Abstract

Proton-translocating Fo∙F1-ATPase/synthase that catalyzes synthesis and hydrolysis of ATP is commonly considered to be a reversibly functioning complex. We have previously shown that venturicidin, a specific Fo-directed inhibitor, blocks the synthesis and hydrolysis of ATP with a significant difference in the affinity [Zharova, T. V. and Vinogradov, A. D. (2017) Biochim. Biophys. Acta, 1858, 939-944]. In this paper, we have studied in detail inhibition of Fo∙F1-ATPase/synthase by venturicidin in tightly coupled membranes of Paracoccus denitrificans under conditions of membrane potential generation. ATP hydrolysis was followed by the ATP-dependent succinate-supported NAD+ reduction (potential-dependent reverse electron transfer) catalyzed by the respiratory chain complex I. It has been demonstrated that membrane energization did not affect the affinity of Fo∙F1-ATPase/synthase for venturicidin. The dependence of the residual ATP synthase activity on the concentration of venturicidin approximated a linear function, whereas the dependence of ATP hydrolysis was sigmoidal: at low inhibitor concentrations venturicidin strongly inhibited ATP synthesis without decrease in the rate of ATP hydrolysis. A model is proposed suggesting that ATP synthesis and ATP hydrolysis are catalyzed by two different forms of Fo∙F1.

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Abbreviations

ΔΨ:

transmembrane electric potential

CCCP:

carbonyl cyanide m-chlorophenylhydrazone

F1 :

hydrophilic part of ATP synthase

Fo :

membrane part of ATP synthase

pmf :

proton motive force (difference between electrochemical potentials of hydrogen ions across a coupling membrane)

S-13:

5-chloro-3-tert-butyl-2′-chloro-4′-nitrosalicylanilide

SBPs:

subbacterial particles of Paracoccus denitrificans

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Acknowledgments

We fondly remember Andrey Dmitrievich Vinogradov, an outstanding scientist and our teacher, who had headed our laboratory and raised multiple generations of scientists, who have worked and currently work in Russia and abroad. We are eternally grateful to him for many years of collaborative work.

We thank Dr. Anna Brzyska and Mr. Grigory Gladyshev for their linguistic help in preparation of this manuscript.

Funding

The work was supported by the Russian Science Foundation (project no. 22-24-00106).

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Biology, Lomonosov Moscow State University, 119234, Moscow, Russia

    Tatyana V. Zharova & Vera G. Grivennikova

  2. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119234, Moscow, Russia

    Vladimir S. Kozlovsky

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  1. Tatyana V. Zharova
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Contributions

T.V.Z. oversaw the study and conducted experiments; T.V.Z., V.G.G., and V.S.K. discussed the results; V.S.K. developed the mathematical model; T.V.Z. and V.G.G. wrote the manuscript.

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Correspondence to Tatyana V. Zharova.

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The authors declare no conflicts of interest in financial or any other sphere. This article does not include the experiments involving humans or animals performed by any of the authors.

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Zharova, T.V., Kozlovsky, V.S. & Grivennikova, V.G. Interaction of Venturicidin and Fo·F1-ATPase/ATP Synthase of Tightly Coupled Subbacterial Particles of Paracoccus denitrificans in Energized Membranes. Biochemistry Moscow 87, 742–751 (2022). https://doi.org/10.1134/S0006297922080065

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