Characterization of 4-hydroxy-2-nonenal-modified peptides by liquid chromatography - Tandem mass spectrometry using data-dependent acquisition: Neutral loss-driven MS 3 versus neutral loss-driven electron capture dissociation

Navin Rauniyar, Stanley M. Stevens, Katalin Prokai-Tatrai, Laszlo Prokai

Research output: Contribution to journalArticle

45 Scopus citations

Abstract

Reactive oxygen species generated during oxidative stress can lead to unfavorable cellular consequences, predominantly due to formation of 4-hydroxy-2-nonenal (HNE) during lipid peroxidation. Data-dependent and neutral loss (NL)-driven MS 3 acquisition have been reported for the identification of HNE adducts by mass spectrometrybased proteomics. However, the limitation associated with this method is the ambiguity in correct assignment of the HNE modification site when more than one candidate site is present as MS 3 is triggered on the neutral loss ion. We introduce NL-triggered electron capture dissociation tandem mass spectrometry (NLECD-MS/MS) for the characterization of HNE-modification sites in peptides. With this method performed using a hybrid linear ion trap-Fourier transform ion cyclotron resonance (FTICR) mass spectrometer, ECD in the FTICR unit of the instrument is initiated on precursor ions of peptides showing the neutral loss of 156 Da corresponding to an HNE molecule in the prescan acquired via collision-induced dissociation tandem mass spectrometry in the linear ion trap. In addition to manifold advantages associated with the ECD method of backbone fragmentation, including extensive sequence fragments, ECD tends to retain the HNE group during MS/MS of the precursor ion, facilitating the correct localization of the modification site. The results also suggest that predisposition of a peptide molecular ion to lose HNE during collision-induced dissociation-based fragmentation is independent of its charge state (2+ or 3+). In addition, we have demonstrated that coupling of solid-phase enrichment of HNE-modified peptides facilitates the detection of this posttranslational modification by NL-driven strategies for low-abundance proteins that are susceptible to substoichiometric carbonylation during oxidative stress.

Original languageEnglish
Pages (from-to)782-789
Number of pages8
JournalAnalytical Chemistry
Volume81
Issue number2
DOIs
StatePublished - 15 Jan 2009

Fingerprint Dive into the research topics of 'Characterization of 4-hydroxy-2-nonenal-modified peptides by liquid chromatography - Tandem mass spectrometry using data-dependent acquisition: Neutral loss-driven MS <sup>3</sup> versus neutral loss-driven electron capture dissociation'. Together they form a unique fingerprint.

  • Cite this