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Physiological role of ATP3-M23K mutation and its impact on yeast mitochondrial ATPase activityтезисы доклада Тезисы
- Авторы: Zubareva Valeria M., Lapashina Anna A., Galkina Kseniia V., Markova Olga V., Sokolov Svyatoslav S., Mamedova Dzhamilia F., Knorre Dmitry A., Feniouk Boris A.
- Сборник: Biochimica et Biophysica Acta (BBA) - Bioenergetics
- Серия: Supplement 2024 EBEC abstracts
- Том: 1865
- Тезисы
- Год издания: 2024
- Издательство: Elsevier BV
- Местоположение издательства: Netherlands
- Номер статьи: 149318
- DOI: 10.1016/j.bbabio.2024.149318
- Аннотация: ATP synthase is a key enzyme in mitochondrial bioenergetics. Typically, it utilizes the energy of transmembrane proton potential difference, maintained by the respiratory chain, to synthesize ATP. However, under membrane de-energization, ATP synthase can hydrolyze ATP. This ATPase activity can significantly decrease intracellular ATP levels and thus requires strict regulation. A common regulatory mechanism is non-competitive inhibition of the enzyme ATPase activity by Mg-ADP (ADP-inhibition) [1]. Certain mutations are known to affect the extent of ADP-inhibition; one of them is γM23K, previously studied in Rhodobacter capsulatus, Bacillus sp. PS3 and Escherichia coli. In these organisms, the M23K mutation has been shown to enhance ADP-inhibition [2,3]. Here, we have obtained a yeast strain with γM23K mutation using the CRISPR-Cas system. Measurements of ATPase activity of isolated mitochondria and submitochondrial particles have shown the increased level of ADP-inhibition. The inhibitory effect of ADP added to the ATPase reaction was more pronounced in samples obtained from the mutant strain in comparison with a parent strain. When ATPase activity was measured in the ATP-regenerating system, the mutant demonstrated a long lag period after adding ATP in the sample, which may also mean that ADP-inhibition in the mutant enzyme is enhanced. On a physiological level, the mutation resulted in a decrease in maximal growth rate (μ) both for Rho+ (with mtDNA) and Rho0 (without mtDNA) cells. However, if we used starved 7-day stationary cultures for inoculation, the mutant Rho0 strain had increased μ in comparison with wild-type Rho0 cells. The work is supported by Russian Science Foundation grant 20-1400268. [1] A.S. Lapashina, B.A. Feniouk, ADP-Inhibition of H+-FF-ATP Synthase, Biochemistry (Moscow), 83 (2018) 1141–1160. [2] S. Bandyopadhyay, W.S. Allison, GammaM23K, gammaM232K, and gammaL77K single substitutions in the TF1-ATPase lower ATPase activity by disrupting a cluster of hydrophobic side chain, Biochemistry, 43 (2004) 9495–9501. [3] B.A. Feniouk, A. Rebecchi, D. Giovannini, S. Anefors, A.Y. Mulkidjanian, W. Junge, et al., Met23Lys mutation in subunit gamma of F(O)F(1)-ATP synthase from Rhodobacter capsulatus impairs the activation of ATP hydrolysis by protonmotive force, Biochim Biophys Acta, 1767 (2007) 1319–1330.
- Добавил в систему: Лапашина Анна Сергеевна