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Technical Brief
Detection and identification of 4-hydroxy-2-nonenal Schiff-base adducts along with products of Michael addition using data-dependent neutral loss-driven MS3 acquisition: Method evaluation through an in vitro study on cytochrome c oxidase modifications
Article first published online: 21 SEP 2009
DOI: 10.1002/pmic.200900116
Copyright © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Additional Information
How to Cite
Rauniyar, N. and Prokai, L. (2009), Detection and identification of 4-hydroxy-2-nonenal Schiff-base adducts along with products of Michael addition using data-dependent neutral loss-driven MS3 acquisition: Method evaluation through an in vitro study on cytochrome c oxidase modifications. Proteomics, 9: 5188–5193. doi: 10.1002/pmic.200900116
Publication History
- Issue published online: 17 NOV 2009
- Article first published online: 21 SEP 2009
- Manuscript Accepted: 11 AUG 2009
- Manuscript Revised: 29 JUL 2009
- Manuscript Received: 20 FEB 2009
Funded by
- National Institutes of Health. Grant Numbers: AG025384, AA015982
- University of North Texas Health Science Center. Grant Number: endowment BK-0031
Keywords:
- 4-Hydroxy-2-nonenal;
- Michael addition;
- Neutral loss-driven MS3;
- Protein adducts;
- Schiff base;
- Technology
Abstract
We report a data-dependent neutral-loss-driven MS3 acquisition to enhance, in addition to abundant Michael adducts, the detection of Schiff-base adducts of proteins and 4-hydroxy-2-nonenal, a reactive end product of lipid peroxidation. In vitro modification of cytochrome c oxidase, a mitochondrial protein complex, was used as a model to evaluate the method. The technique allowed for a confident validation of modification sites and also identified a Schiff-base adduct in subunit Vb of the protein complex.