Enzymatic Oxidation of Polyunsaturated Fatty Acids

Nicolaou, Anna; Murphy, SA; Al-Aaswad, Naser; Spickett, Corinne M.; Forman, Henry Jay

doi:10.1201/b18138-8

Cited by 1 publication

(1 citation statement)

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“…Some of the peptides identified contained missed cleavages owing to modification of lysine residues, which could complicate relative quantification against control samples, but all showed the expected diagnostic ions. A number of the lysines and histidines observed to be modified were ones that have been reported previously to be susceptible to electrophilic attack, and the occurrence of acrolein-protein adducts has previously been reported both in vitro and in vivo [22,23], including studies of acrolein modification of albumin [24]. It is also worth noting that in a biological sample the product of a fully reduced malondialdehyde adduct, which could be formed from either lipoxidation and glucoxidation events, would be indistinguishable from the acrolein adduct.…”

Section: Accepted Manuscriptmentioning

confidence: 77%

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

Afonso

Sousa

Pitt

et al. 2018

Archives of Biochemistry and Biophysics

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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 NaBH or NaBHCN 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 adducts, and the diagnostic ions will facilitate the development of targeted MS methods for analysis of adducts in more complex samples.

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Section: Accepted Manuscriptmentioning

confidence: 77%