Journal
MOLECULAR NUTRITION & FOOD RESEARCH
Volume 55, Issue 9, Pages 1401-1410Publisher
WILEY
DOI: 10.1002/mnfr.201100255
Keywords
Acrolein; Aldehyde/keto-reactive probe; Mitochondria; Protein carbonyls; Selected reaction monitoring
Categories
Funding
- NIH [R01 AG025372, S10 RR022589, R01 HL081721]
- Environmental Health Sciences Center [P30 ES00210]
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Acrolein (ACR) exposure leads to the formation of protein-ACR adducts. Protein modification by ACR has been associated with various chronic diseases including cardiovascular and neurodegenerative diseases. Here, we report an analytical strategy that enables the quantification of Michael-type protein adducts of ACR in mitochondrial proteome samples using liquid chromatography in combination with tandem mass spectrometry and selected ion monitoring (LC-MS/MS SRM) analysis. Our approach combines site-specific identification and relative quantification at the peptide level of protein-ACR adducts in relation to the unmodified protein thiol pool. Treatment of 3-month-old rats with CCl4, an established in vivo model of acute oxidative stress, resulted in significant increases in the ratios of distinct ACR-adducted peptides to the corresponding unmodified thiol-peptides obtained from proteins that were isolated from cardiac mitochondria. The mitochondrial proteins that were found adducted by ACR were malate dehydrogenase, NADH dehydrogenase [ubiquinone] flavoprotein 1, cytochrome c oxidase subunit VIb isoform 1, ATP synthase d chain, and ADP/ATP translocase 1. The findings indicate that protein modification by ACR has potential value as an index of mitochondrial oxidative stress.
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