Protein Methionine Sulfoxide Dynamics in Arabidopsis thaliana under Oxidative Stress
Silke Jacques, Bart Ghesquiere, Pieter-Jan De Bock, Hans Demol, Khadija Wahn, Patrick Willems, Joris Messens, Frank Van Breusegem and Kris Gevaert
Molecular & Cellular Proteomics (2015) 14: 1217-1229
This work investigates methionine oxidation, which is a reversible post-translational modification and a mechanism by which proteins perceive oxidative stress. COFRADIC technology was used to specifically isolate peptides carrying in vivo oxidized methionine.
Arabidopsis catalase 2 knock-out and wild type plants were oxidized by increased concentrations of hydrogen peroxide. The proteins were extracted, desalted, and digested. The peptides were then fractionated and labelled. Those originating from cat-2 knockout were 12C3 propionated, whereas the wild type was 13C3 propionated. LC/MS/MS analysis and database searching resulted in the identification of 513 oxidation sites in 403 proteins.
Quantification of methionine oxidation using Mascot Distiller revealed 55 MetOx peptides in 51 proteins that were significantly more oxidized in the catalase knock-out. The data have been uploaded into the PRIDE database, project accession number PXD001286. |