Redox proteomic identification of HNE-bound mitochondrial proteins in cardiac tissues reveals a systemic effect on energy metabolism after doxorubicin treatment

• Published in: Free radical biology & medicine Vol. 72; pp. 55 - 65
• Main Authors: Zhao, Y., Miriyala, S., Miao, L., Mitov, M., Schnell, D., Dhar, S.K., Cai, J., Klein, J.B., Sultana, R., Butterfield, D.A., Vore, M., Batinic-Haberle, I., Bondada, S., St. Clair, D.K.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2014
• Subjects: Aldehydes - metabolism; Animals; Antibiotics, Antineoplastic - toxicity; ATP synthase; Cardiac injury; Dihydrolipoyl dehydrogenase; Doxorubicin; Doxorubicin - toxicity; Electrophoresis, Gel, Two-Dimensional; Energy Metabolism - drug effects; Free radicals; Immunoblotting; Immunoprecipitation; Lipid Peroxidation - drug effects; Male; Metabolism; Mice; Mice, Inbred C57BL; Mitochondria, Heart - drug effects; Mitochondria, Heart - metabolism; NADH dehydrogenase [ubiquinone] iron–sulfur protein 2; Oxidation-Reduction; Oxidative stress; Proteomics; Redox proteomics; Succinate dehydrogenase [ubiquinone] flavoprotein; Oxidative stress; FH; Doxorubicin; CKMT2; ATP5B; ATP synthase; Succinate dehydrogenase [ubiquinone] flavoprotein; Free radicals; Cardiac injury; TCA cycle; Oxct1; Dihydrolipoyl dehydrogenase; DOX; SDHA; Metabolism; MnP; ECAR; HADHA; NAC; Redox proteomics; NADH dehydrogenase [ubiquinone] iron–sulfur protein 2; ETC; ROS; NDUFS2; DLD; HNE; OCR
• ISSN: 0891-5849; 1873-4596
• DOI: 10.1016/j.freeradbiomed.2014.03.001

Doxorubicin (DOX), one of the most effective anticancer drugs, is known to generate progressive cardiac damage, which is due, in part, to DOX-induced reactive oxygen species (ROS). The elevated ROS often induce oxidative protein modifications that result in alteration of protein functions. This study demonstrates that the level of proteins adducted by 4-hydroxy-2-nonenal (HNE), a lipid peroxidation product, is significantly increased in mouse heart mitochondria after DOX treatment. A redox proteomics method involving two-dimensional electrophoresis followed by mass spectrometry and investigation of protein databases identified several HNE-modified mitochondrial proteins, which were verified by HNE-specific immunoprecipitation in cardiac mitochondria from the DOX-treated mice. The majority of the identified proteins are related to mitochondrial energy metabolism. These include proteins in the citric acid cycle and electron transport chain. The enzymatic activities of the HNE-adducted proteins were significantly reduced in DOX-treated mice. Consistent with the decline in the function of the HNE-adducted proteins, the respiratory function of cardiac mitochondria as determined by oxygen consumption rate was also significantly reduced after DOX treatment. Treatment with Mn(III) meso-tetrakis(N-n-butoxyethylpyridinium-2-yl)porphyrin, an SOD mimic, averted the doxorubicin-induced mitochondrial dysfunctions as well as the HNE–protein adductions. Together, the results demonstrate that free radical-mediated alteration of energy metabolism is an important mechanism mediating DOX-induced cardiac injury, suggesting that metabolic intervention may represent a novel approach to preventing cardiac injury after chemotherapy. •Doxorubicin increased total HNE-adducted proteins in cardiac mitochondria.•Redox proteomics identified several HNE-targeted cardiac mitochondria proteins important for energy metabolism.•Doxorubicin reduced the activities of complex I, SDHA, ATP synthase, and DLD in ETC.•MnP averted doxorubicin-induced mitochondrial dysfunction and HNE adductions.