Related Experiment Video
Updated: Jul 19, 2026
Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
Published on: June 21, 2021
Redox proteomic identification of oxidized cardiac proteins in adriamycin-treated mice
Yumin Chen1, Chotiros Daosukho, Wycliffe O Opii
1Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40506, USA.
Abstract:
Adriamycin (ADR) is a potent anticancer drug, but its use is limited by a dose-dependent cardiotoxicity. Oxidative stress is regarded as the mediating mechanism of ADR cardiotoxicity. However, cardiac proteins that are oxidatively modified have not been well characterized. We took a redox proteomics approach to identify increasingly oxidized murine cardiac proteins after a single injection of ADR (ip, 20 mg/kg body wt). The specific carbonyl levels of three proteins were significantly increased, and these proteins were identified as triose phosphate isomerase (TPI), beta-enolase, and electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO). TPI and enolase are key enzymes in the glycolytic pathway, and ETF-QO serves as the transporter for electrons derived from a variety of oxidative processes to the mitochondria respiratory chain. Cardiac enolase activity in ADR-treated mice was reduced by 25%, whereas the cardiac TPI activity remained unchanged. Oxidation of purified enolase or TPI via Fenton chemistry led to a 17 or 23% loss of activity, respectively, confirming that a loss of activity was the consequence of oxidation. The observation that these cardiac enzymes involved in energy production are more oxidized resulting from ADR treatment indicates that the bioenergetic pathway is an important target in ADR-initiated oxidative stress.
Insights
Adriamycin (ADR) chemotherapy causes heart damage by oxidizing key energy-producing proteins like beta-enolase and triose phosphate isomerase (TPI). This study identifies these proteins, revealing a new target for preventing ADR cardiotoxicity.
Area of Science:
- Biochemistry
- Cardiology
- Proteomics
Background:
- Adriamycin (ADR) is an effective anticancer drug, but its clinical use is restricted by dose-dependent cardiotoxicity.
- Oxidative stress is the primary mechanism behind ADR-induced cardiotoxicity.
- Specific cardiac proteins oxidized by ADR remain poorly characterized.
Purpose of the Study:
- To identify cardiac proteins that undergo oxidative modification following ADR treatment using a redox proteomics approach.
- To investigate the functional consequences of ADR-induced protein oxidation on cardiac enzymes involved in energy metabolism.
Main Methods:
- A redox proteomics strategy was employed to analyze cardiac protein oxidation in mice after a single ADR injection.
- Specific carbonyl levels were quantified, and affected proteins were identified.
- Enzyme activity assays were performed on cardiac extracts and purified proteins.
Main Results:
- Triose phosphate isomerase (TPI), beta-enolase, and electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO) were identified as significantly oxidized proteins.
- Cardiac enolase activity decreased by 25% in ADR-treated mice.
- Oxidation of purified enolase and TPI resulted in a 17% and 23% loss of activity, respectively.
Conclusions:
- ADR treatment leads to the oxidation and functional impairment of key glycolytic enzymes (TPI, enolase) and mitochondrial electron transporters (ETF-QO) in the heart.
- These findings highlight the cardiac bioenergetic pathway as a critical target for ADR-induced oxidative stress.
- Understanding these specific protein modifications may lead to strategies for mitigating ADR cardiotoxicity.