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. 2020 Sep 1;8:822. doi: 10.3389/fcell.2020.00822

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Copyright © 2020 Xu, Gan, Gao, Huang, Wu, Zhang, Wang and Sheng.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Caffeine has no antioxidant activity in vitro but enhances SIRT3 activity. (A) The structure of caffeine. (B) Caffeine showed no DPPH (free radical) scavenging activity, as evidenced by the absence of any change in absorbance. (C) Caffeine does not directly enhance SOD2 activity in vitro. (D) Caffeine directly enhances SIRT3 activity in an in vitro system. (E) Caffeine promotes SIRT3-mediated deacetylation of SOD2 in vitro. (F) Caffeine directly binds to SIRT3 with high affinity (K_D = 6.562 × 10^{– 7} M). BLI sensorgrams indicating the interactions between gradient concentrations of caffeine and SIRT3 were measured on an Octet Red 96 system, with association and dissociation for 300 s each. Data are expressed as the mean ± SEM of three independent experiments. **P < 0.01, ***P < 0.001 vs. the control.