Journal of the American Society for Mass Spectrometry

, Volume 14, Issue 3, pp 215–226

Study of protein modification by 4-hydroxy-2-nonenal and other short chain aldehydes analyzed by electrospray ionization tandem mass spectrometry


  • François Fenaille
    • Department of Quality and Safety AssuranceNestlé Research Center, Nestec Ltd.
    • Department of Quality and Safety AssuranceNestlé Research Center, Nestec Ltd.
  • Jean-Claude Tabet
    • Laboratoire de Chimie Structurale Organique et BiologiqueUniversity Pierre and Marie Curie

DOI: 10.1016/S1044-0305(02)00911-X

Cite this article as:
Fenaille, F., Guy, P.A. & Tabet, J. J Am Soc Mass Spectrom (2003) 14: 215. doi:10.1016/S1044-0305(02)00911-X


A convenient way to study lipid oxidation products-modified proteins by means of suitable model systems has been investigated. As a model peptide, the oxidized B chain of insulin has been chemically modified by either 4-hydroxy-2-nonenal (HNE) or hexanal and the extent, sites, and structure of modifications were assessed by electrospray mass spectrometry. A reduction step, using either NaCNBH3 or NaBH4, was also studied to stabilize the alkylated compounds. From the data gathered, it appeared that NaCNBH3, when added at the beginning of incubation, dramatically influenced the HNE-induced modifications in terms of the addition mechanism (Schiff base formation instead of Michael addition) but also of the amino acid residues modified (N-terminal amino acid instead of histidine residues). However, by reducing the HNE-adducted species at the end of the reaction with NaBH4, the fragment ions obtained in the product ion scan experiments become more stable and thus, easier to interpret in terms of origin and mechanism involved. With regard to hexanal induced modifications, we have observed that hexanal addition under reductive conditions led to an extensive modification of the peptide backbone. Moreover, as confirmed by “in-source” collision followed by collision induced dissociation (CID) experiments on selected precursor ions (pseudo-MS3 experiments), N,N-di-alkylations were first observed on the N-terminal residue and further on Lys29 residue. On the other hand, compared to the native peptide, no significant changes in MS/MS fragmentation patterns (b and y ions series) were observed whatever the basic site modified by the aldehyde-addition.

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© American Society for Mass Spectrometry 2003