A Comprehensive Analytical Strategy To Identify Malondialdehyde-Modified Proteins and Peptides.

Abstract

Mass spectrometric-based proteomics is a powerful tool to analyze post-translationally modified proteins. Carbonylation modifications that result from oxidative lipid breakdown are a class of post-translational modifications that are poorly characterized with respect to protein targets and function. This is partly due to the lack of dedicated mass spectrometry-based technologies to facilitate the analysis of these modifications. Here, we present a comprehensive approach to identify malondialdehyde-modified proteins and peptides. Malondialdehyde is among the most abundant of the lipid peroxidation products; and malondialdehyde-derived adducts on proteins have been implicated in cardiovascular diseases, neurodegenerative disorders, and other clinical conditions. Our integrated approach targets three levels of the overall proteomic workflow: (i) sample preparation, by employing a targeted enrichment strategy; (ii) high-performance liquid chromatography, by using a gradient optimized for the separation of the modified peptides; and (iii) tandem mass spectrometry, by improving the spectral quality of very low-abundance peptides. By applying the optimized procedure to a whole cell lysate spiked with a low amount of malondialdehyde-modified proteins, we were able to identify up to 350 different modified peptides and localize the modification to a specific lysine residue. This methodology allows the comprehensive analysis of malondialdehyde-modified proteins.

• Authors: Juliane Weißer, Claudia Ctortecka, Clara J Busch, Shane R Austin, Karin Nowikovsky, Koji Uchida, Christoph J Binder, Keiryn L Bennett
• Journal: Analytical chemistry (Anal Chem) Vol. 89 Issue 7 Pg. 3847-3852 (04 04 2017) ISSN: 1520-6882 [Electronic] United States
• PMID: 28248083 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Peptides
• Proteins
• Malondialdehyde

Topics

• Malondialdehyde (analysis)
• Mass Spectrometry
• Molecular Structure
• Peptides (chemistry)
• Proteins (chemistry)