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

Y. Zhao, S. Miriyala, L. Miao, M. Mitov, D. Schnell, S. K. Dhar, J. Cai, J. B. Klein, R. Sultana, D. A. Butterfield, M. Vore, I. Batinic-Haberle, S. Bondada, D. K. St. Clair

Research output: Contribution to journalArticlepeer-review

81 Scopus citations

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.

Original languageEnglish
Pages (from-to)55-65
Number of pages11
JournalFree Radical Biology and Medicine
Volume72
DOIs
StatePublished - Jul 2014

Bibliographical note

Funding Information:
This work is supported by NIH Grants CA 049797 and CA 139843 , Cancer Center Support Grant (CCSG) P30CA177558 from the NCI , and the Edward P. Evans Foundation . The authors thank the University of Kentucky Free Radical Biology in Cancer Shared Resource Facility for its support. We also thank Dr. Liang-Jun Yan of the University of North Texas Health Science Center for the kind gift of synthesized dihydrolipoamide, as well as for his excellent technical help.

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

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

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