title: Redox proteomic identification of HNE-bound mitochondrial proteins in cardiac tissues reveals a systemic effect on energy metabolism after doxorubicin treatment.
abstract: 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.
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA; Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences, Shreveport, LA 71130, USA.
authors.forename: S
authors.initials: S
authors.lastname: Miriyala
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: L
authors.initials: L
authors.lastname: Miao
authors.affiliation: Free Radical Biology in Cancer Shared Resource Facility, Markey Cancer Center, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: M
authors.initials: M
authors.lastname: Mitov
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: D
authors.initials: D
authors.lastname: Schnell
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: S K
authors.initials: SK
authors.lastname: Dhar
authors.affiliation: Department of Nephrology and Proteomics Facility, University of Louisville, Louisville, KY 40292, USA.
authors.forename: J
authors.initials: J
authors.lastname: Cai
authors.affiliation: Department of Nephrology and Proteomics Facility, University of Louisville, Louisville, KY 40292, USA.
authors.forename: J B
authors.initials: JB
authors.lastname: Klein
authors.affiliation: Department of Chemistry, Center of Membrane Sciences, and Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: R
authors.initials: R
authors.lastname: Sultana
authors.affiliation: Free Radical Biology in Cancer Shared Resource Facility, Markey Cancer Center, University of Kentucky, Lexington, KY 40506, USA; Department of Chemistry, Center of Membrane Sciences, and Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Butterfield
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: M
authors.initials: M
authors.lastname: Vore
authors.affiliation: Department of Radiation Oncology, Duke University School of Medicine, Durham, NC 27710, USA.
authors.forename: I
authors.initials: I
authors.lastname: Batinic-Haberle
authors.affiliation: Department of Immunology, University of Kentucky, Lexington, KY 40506, USA.
authors.forename: S
authors.initials: S
authors.lastname: Bondada
authors.affiliation: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA. Electronic address: dstcl00@uky.edu.
authors.forename: D K
authors.initials: DK
authors.lastname: St Clair
mesh_terms: D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D004317:Doxorubicin / Q000633:toxicity*; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism / Q000187:drug effects*; D015151:Immunoblotting; D047468:Immunoprecipitation; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000378:metabolism; D010084:Oxidation-Reduction; D040901:Proteomics
chemical_list: D000447:Aldehydes; D000903:Antibiotics, Antineoplastic; C462359:tert-4-hydroxy-2-nonenal; D004317:Doxorubicin
keywords: ATP synthase; Cardiac injury; Dihydrolipoyl dehydrogenase; Doxorubicin; Free radicals; Metabolism; NADH dehydrogenase [ubiquinone] iron–sulfur protein 2; Oxidative stress; Redox proteomics; Succinate dehydrogenase [ubiquinone] flavoprotein