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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.