Abstract
Na,K-ATPase is a crucial enzyme responsible for maintaining Na+, K+-gradients across the cell membrane, which is essential for numerous physiological processes within various organs and tissues. Due to its significance in cellular physiology, inhibiting Na,K-ATPase can have profound physiological consequences. This characteristic makes it a target for various pharmacological applications, and drugs that modulate the pump’s activity are thus used in the treatment of several medical conditions. Cytochrome c (Cytc) is a protein with dual functions in the cell. In the mitochondria, it is essential for ATP synthesis and energy production. However, in response to apoptotic stimuli, it is released into the cytosol, where it triggers programmed cell death through the intrinsic apoptosis pathway. Aside from its role in canonical intrinsic apoptosis, Cytc also plays additional roles. For instance, Cytc participates in certain non-apoptotic functions –those which are less well-understood in comparison to its role in apoptosis. Within this in vitro study, we have shown the impact of Cytc on Na,K-ATPase for the first time. Cytc has a biphasic action on Na,K-ATPase, with activation at low concentrations (0.06 ng/ml; 6 ng/ml) and inhibition at high concentration (120 ng/ml). Cytc moreover displays isoform/subunit specificity and regulates the Na+ form of the enzyme, while having no effect on the activity or kinetic parameters of the K+-dependent form of the enzyme. Changing the affinity of p-chloromercuribenzoic acid (PCMB) by Cytc is therefore both a required and sufficient condition for confirming that PCMB and Cytc share the same target, namely the thiol groups of cysteine in Na,K-ATPase.
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Gvantsa Chkadua—performed the experiments; formulated hypothesis; developed the study objectives; defined experimental, statistical, and analytical approaches; analyzed the data; researched the literature; and wrote the paper.
Eka Nozadze—Performed the experiments, theoretical calculation, and data analysis (statistical or other), and researched the literature.
Leila Tsakadze—performed the experiments, and contributed to acquisition, analysis and interpretation of the data.
Lia Shioshvili—Performed the experiments; and contributed to acquisition, analysis and interpretation of the data.
Nana Arutinova—performed the experiments, and collected and analyzed the data.
Marine Leladze—performed the experiments, and collected and analyzed the data.
Sopio Dzneladze—performed the experiments, and collected and analyzed the data.
Maia Javakhishvili—performed the experiments, and collected and analyzed the data.
Tamar Jariashvili—performed the experiments, and collected and analyzed the data.
Elene Petriashvili—performed the experiments, and collected and analyzed the data.
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Chkadua, G., Nozadze, E., Tsakadze, L. et al. The effect of cytochrome c on Na,K-ATPase. J Bioenerg Biomembr 56, 221–234 (2024). https://doi.org/10.1007/s10863-024-10012-3
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DOI: https://doi.org/10.1007/s10863-024-10012-3