Revisiting the pro-oxidant activity of copper: interplay of ascorbate, cysteine, and glutathione

Falcone, Enrico; Stellato, Francesco; Vileno, Bertrand; Bouraguba, Merwan; Lebrun, Vincent; Ilbert, Marianne; Morante, Silvia; Faller, Peter

Abstract

Copper (Cu) is essential for most organisms, but it can be poisonous in excess, through mechanisms such as protein aggregation, trans-metallation, and oxidative stress. The latter could implicate the formation of potentially harmful reactive oxygen species (O2, H2O2, and HO) via the redox cycling between Cu(II)/Cu(I) states in the presence of dioxygen and physiological reducing agents such as ascorbate (AscH), cysteine (Cys), and the tripeptide glutathione (GSH). Although the reactivity of Cu with these reductants has been previously investigated, the reactions taking place in a more physiologically relevant mixture of these biomolecules are not known. Hence, we report here on the reactivity of Cu with binary and ternary mixtures of AscH, Cys, and GSH. By measuring AscH and thiol oxidation, as well as HO formation, we show that Cu reacts preferentially with GSH and Cys, halting AscH oxidation and also HO release. This could be explained by the formation of Cu-thiolate clusters with both GSH and, as we first demonstrate here, Cys. Moreover, we observed a remarkable acceleration of Cu-catalyzed GSH oxidation in the presence of Cys. We provide evidence that both thiol-disulfide exchange and the generated H2O2 contribute to this effect. Based on these findings, we speculate that Cu-induced oxidative stress may be mainly driven by GSH depletion and/or protein disulfide formation rather than by HO and envision a synergistic effect of Cys on Cu toxicity.

Citation

Falcone, E., Stellato, F., Vileno, B., Bouraguba, M., Lebrun, V., Ilbert, M., Morante, S., & Faller, P. (2023). Revisiting the pro-oxidant activity of copper: interplay of ascorbate, cysteine, and glutathione. Metallomics, 15(7), Article mfad040. https://doi.org/10.1093/mtomcs/mfad040

• Journal Article Type: Article
• Acceptance Date: Jun 7, 2023
• Online Publication Date: Jun 23, 2023
• Publication Date: Jul 10, 2023
• Deposit Date: Jun 29, 2023
• Publicly Available Date: Jun 29, 2023
• Journal: Metallomics
• Print ISSN: 1756-5901
• Electronic ISSN: 1756-591X
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 15
• Issue: 7
• Article Number: mfad040
• DOI: https://doi.org/10.1093/mtomcs/mfad040
• Keywords: Metals and Alloys; Biochemistry; Biomaterials; Biophysics; Chemistry (miscellaneous)
• Public URL: https://nottingham-repository.worktribe.com/output/22436669
• Publisher URL: https://academic.oup.com/metallomics/advance-article/doi/10.1093/mtomcs/mfad040/7206412