A mass spectrometry approach for the identification and localization of small aldehyde modifications of proteins

Catarina B. Afonso, Bebiana C. Sousa, Andrew R. Pitt, Corinne M. Spickett

Research output: Contribution to journalArticle

Abstract

Lipids containing polyunsaturated fatty acids are primary targets of oxidation, which produces reactive short-chain aldehydes that can covalently modify proteins, a process called lipoxidation. Improved mass spectrometry (MS) methods for the analysis of these adducts in complex biological systems are needed. Lysozyme and human serum albumin (HSA) were used as model proteins to investigate lipoxidation products formed by two short-chain aldehydes, acrolein and pentanal, which are unsaturated and saturated aldehydes respectively. The adducts formed were stabilized by NaBH4 or NaBH3CN reduction and analysed by MS. Analysis of intact modified lysozyme showed a pentanal modification resulting from Schiff's base formation (+70 Da), and up to 8 acrolein adducts, all resulting from Michael addition (+58 Da). Analysis of tryptic digests identified specific histidine, cysteine and lysine residues modified in both lysozyme and HSA, and determined characteristic amino acid-specific fragmentations. Eight different internal fragment ions were found that could be used as general diagnostic ions for pentanal- and acrolein-modified amino acids. The combined use of intact protein analysis and LC-MS/MS methods provided a powerful tool for the identification and localization of aldehyde-protein adduct, and the diagnostic ions will facilitate the development of targeted MS methods for analysis of adducts in more complex samples.

Original languageEnglish
Pages (from-to)38-45
JournalArchives of Biochemistry and Biophysics
Volume646
Early online date23 Mar 2018
DOIs
Publication statusPublished - 15 May 2018

Bibliographical note

© 2018, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International

Funding: European Union's Horizon 2020 research and innovation programme under the Marie Sklowdowska-Curie grant agreement number 675132

Keywords

  • Acrolein
  • Lipoxidation;
  • Pentanal
  • Diagnostic ions
  • Aldehydes
  • Immonium ion

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