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Antibiotic Pyrrolomycin as an Efficient Mitochondrial Uncoupler

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Abstract

Pyrrolomycins C (Pyr_C) and D (Pyr_D) are antibiotics produced by Actinosporangium and Streptomyces. The mechanism of their antimicrobial activity consists in depolarization of bacterial membrane, leading to the suppression of bacterial bioenergetics through the uncoupling of oxidative phosphorylation, which is based on the protonophore action of these antibiotics [Valderrama et al., Antimicrob. Agents Chemother. (2019) 63, e01450]. Here, we studied the effect of pyrrolomycins on the isolated rat liver mitochondria. Pyr_C was found to be more active than Pyr_D and uncoupled mitochondria in the submicromolar concentration range, which was observed as the mitochondrial membrane depolarization and stimulation of mitochondrial respiration. In the case of mitoplasts (isolated mitochondria with impaired outer membrane integrity), the difference in the action of Pyr_C and Pyr_D was significantly less pronounced. By contrast, in inverted submitochondrial particles (SMPs), Pyr_D was more active as an uncoupler, which caused collapse of the membrane potential even at the nanomolar concentrations. The same ratio of the protonophoric activity of Pyr_D and Pyr_C was obtained by us on liposomes loaded with the pH indicator pyranine. The protonophore activity of Pyr_D in the planar bilayer lipid membranes (BLMs) was maximal at ~pH 9, i.e., at pH values close to pKa of this compound. Pyr_D functions as a typical anionic protonophore; its activity in the BLM could be reduced by the addition of the dipole modifier phloretin. The difference between the protonophore activity of Pyr_C and Pyr_D in the mitochondria and BLMs can be attributed to a higher ability of Pyr_C to penetrate the outer mitochondrial membrane.

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Abbreviations

ACMA:

9-amino-6-chloro-2-methoxyacridine

BLM:

bilayer lipid membrane

CCCP:

carbonyl cyanide-m-chlorophenyl hydrazone

DNP:

2,4-dinitrophenol

Pyr_C:

Pyrrolomycin C

Pyr_D:

pyrrolomycin D

SMP:

submitochondrial particles

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Acknowledgments

The authors are grateful to Dr. Ruben C. Hartkoorn (University of Lille, France) for kindly providing pyrrolomycins, I. K. Gorelova (Faculty of Bioengineering and Bioinformatics, Moscow State University) for helping with the experiments in the BLMs, and V. G. Grivennikova (Department of Biochemistry, Faculty of Biology, Lomonosov Moscow State University) for kindly providing the SMP preparations.

Funding

The study was supported by the Russian Science Foundation (project no. 21-14-00062).

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Authors and Affiliations

  1. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    Alexander M. Firsov, Ljudmila S. Khailova, Tatyana I. Rokitskaya, Elena A. Kotova & Yuri N. Antonenko

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  1. Alexander M. Firsov
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  2. Ljudmila S. Khailova
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  3. Tatyana I. Rokitskaya
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  4. Elena A. Kotova
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Contributions

Y. N. Antonenko developed the concept and managed the study; A. M. Firsov, L. S. Khailova, and T. I. Rokitskaya conducted the experiments; T. I. Rokitskaya, E. A. Kotova, and Y. N. Antonenko discussed the results; E. A. Kotova and Y. N. Antonenko wrote and edited the manuscript.

Corresponding author

Correspondence to Yuri N. Antonenko.

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The authors declare no conflicts of interest. The article does not contain description of studies involving humans or animals performed by any of the authors.

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Firsov, A.M., Khailova, L.S., Rokitskaya, T.I. et al. Antibiotic Pyrrolomycin as an Efficient Mitochondrial Uncoupler. Biochemistry Moscow 87, 812–822 (2022). https://doi.org/10.1134/S0006297922080120

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  • DOI: https://doi.org/10.1134/S0006297922080120

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