Advertisement

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

Authors

  • Navin Rauniyar,

    1. Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, TX, USA
    Search for more papers by this author
  • Laszlo Prokai

    Corresponding author
    1. Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, TX, USA
    • Department of Molecular Biology & Immunology, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107, USA Fax: +1-817-735-2118
    Search for more papers by this author

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.

Ancillary