Different Detergent Stability of ATP Synthase from Pea, Winter Wheat and Maize Mitochondria
Author(s)
I. V. Ukolova, M. A. Kondakova, G. B. Borovskii
Abstract
Mitochondrial ATP synthase is a macromolecular nanomachine, which produces most of the ATP in the cell. This enzyme is located in inner mitochondrial membrane in the form of dimers, which assemble into long rows at the cristae rims. Dimers of ATP synthase are sensitive to detergent treatment in many organisms, as a result of which they mainly dissociate into monomers. Plant enzyme is also very detergent-sensitive, nevertheless, it can be assumed that the detergent-sensitivity or, vice versa, detergent-stability of the enzyme under the treat-ment may vary in different plant species. In this regard, the aim of this work was to study the detergent-stability, as well as the activity of various forms of ATP synthase, solubilized from the mitochondria of different plant species. For this purpose, we used organelles isolated from etiolated pea, winter wheat, and maize shoots. These species belong to different families (Poaceae and Fabaceae), clades (monocots and dicots) and, in addition, differ in their low temperature tolerance. For the solubilization of organelles, a mild non-ionic detergent digitonin was used, which preserved and stabilized the supramolecular associations of membrane proteins. Using 1D BN-PAGE followed by ingel enzyme activity assay, it was shown that ATP synthase in all studied species was solubilized mainly as monomers Va and Vb, dimer Vb2, supercomplex IV1Va2, and minor subcomplex F1. In the course of the study, for the first time, differences in the detergent-stability of dimeric and monomeric forms of the enzyme between the studied species were revealed. It was found that the dimeric form in maize and the monomeric form in winter wheat were the most stable; while, pea ATP synthase had the highest activity. The relationship between the revealed features and the life strategy of the species is assumed and discussed.
About the Authors
Ukolova Irina Vladimirovna, Candidate of Science (Biology), Senior Researcher, Siberian Institute of Plant Physiology and Biochemistry SB RAS, 132, Lermontov st., Irkutsk, 664033, Russian Federation, e-mail: irina@sifibr.irk.ru
Kondakova Marina Alexandrovna, Candidate of Science (Biology), Leading Engineer, Siberian Institute of Plant Physiology and Biochemistry SB RAS, 132, Lermontov st., Irkutsk, 664033, Russian Federation, e-mail: kondakova-marina@mail.ru
Borovskii Gennadii Borisovich, Doctor of Sciences (Biology), Professor, Principal Research Scientist, Siberian Institute of Plant Physiology and Biochemistry SB RAS, 132, Lermontov st., Irkutsk, 664033, Russian Federation, e-mail: borovskii@sifibr.irk.ru
For citation
Ukolova I.V., Kondakova M.A., Borovskii G.B. Different Detergent Stability of ATP Synthase from Pea, Winter Wheat and Maize Mitochondria. The Bulletin of Irkutsk State University. Series Biology. Ecology, 2021, vol. 37, pp. 111-116. https://doi.org/10.26516/2073-3372.2021.37.111 (in Russian)
Keywords
ATP synthase, BN-PAGE, in-gel activity assay, digitonin, pea, winter wheat, maize.
UDC
577.22:577.23
DOI
https://doi.org/10.26516/2073-3372.2021.37.111
References
Dudkina N.V., Sunderhaus S., Braun H.P., Boekema E.J. Characterization of dimeric ATP synthase and cristae membrane ultrastructure from Saccharomyces and Polytomella mitochondria. FEBS Lett., 2006, vol. 580, no. 14, pp. 3427-3432. https://doi.org/10.1016/j.febslet.2006.04.097
Kühlbrandt W. Structure and mechanisms of F-type ATP synthases. Annu Rev. Biochem. 2019, vol. 88, pp. 515-549. https://doi.org/10.1146/annurev-biochem-013118-110903
Ukolova I.V., Kondakova M.A., Kondratov I.G., Sidorov A.V., Borovskii G.B., Voinikov V.K. New insights into the organisation of the oxidative phosphorylation system in the example of pea shoot mitochondria. Biochim. Biophys. Acta Bioenerg., 2020, vol. 1861, no. 11, art. 148264. https://doi.org/10.1016/j.bbabio.2020.148264
Miranda-Astudillo H., Colina-Tenorio L., Jiménez-Suárez A., Vázquez-Acevedo M., Salin B., Giraud M. F., Remacle C., Cardol P., González-Halphen D. Oxidative phosphorylation supercomplexes and respirasome reconstitution of the colorless alga Polytomella sp. Biochim. Biophys. Acta Bioenerg., 2018, vol. 1859, no. 6, pp. 434-444. https://doi.org/10.1016/j.bbabio.2018.03.004
Zancani M., Braidot E., Filippi A., Lippe G. Structural and functional properties of plant mitochondrial F-ATP synthase. Mitochondrion, 2020, vol. 53, pp. 178-193. https://doi.org/10.1016/j.mito.2020.06.001
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