The detection and comparison of the amount of superoxide anion (O2.-) released by different complexes in mitochondrial electron transport chain (ETC) can locate the main electron leakage sites in mitochondria. In order to realize this, we designed an ultrasensitive photoelectrochemical (PEC) sensor by in situ hydrothermal growth of MnO2 nanosheets on Co3O4 nanowires array modified Ti substrate (NWA|Ti). Due to the formation of a core-shell p-n heterojunction with high specific surface area, tight surface contact and plentiful oxygen vacancies (OVs), MnO2@Co3O4 NWA|Ti possesses a strong visible light absorption, high charges transfer and separation ability. The proposed PEC sensor exhibited a wide linear range of 0.1-50000 nM and a low detection limit of 0.025 nM towards H2O2. Due to the rapid conversion of O2.- to H2O2 inside mitochondria, the PEC sensor can indirectly monitor the electron leakage in the ETC. Specifically, four selected mitochondrial inhibitors specifically inhibited the corresponding complex in mitochondria extracted from living HepG2 cells (hepatocellular carcinoma cells), and the H2O2 levels converted from O2.- was measured by the PEC sensor. It is evident that IQ (ubiquinone binding) site of complex I and Qo (ubiquinol oxidation) site of complex III are the key sites at which electron leakage occurred. This study could provide meaningful information for the diagnosis and treatment of certain disease caused by oxidative stress due to the electron leakage.
Keywords: Core-shell p-n heterojunction; HepG2 cells; Mitochondrial inhibitors; Photoelectrochemical sensor; Superoxide anion.
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