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.
We report a data-dependent neutral-loss-driven MS(3) 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.
Rauniyar
Navin
N
Department of Molecular Biology and Immunology, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107, USA.
Prokai
Laszlo
L
D000447:Aldehydes / Q000737:chemistry*; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D015203:Reproducibility of Results; D012545:Schiff Bases / Q000737:chemistry*
D000447:Aldehydes; D011506:Proteins; D012545:Schiff Bases; D003576:Electron Transport Complex IV; C027576:4-hydroxy-2-nonenal