Peptide-phospholipid cross-linking reactions: identification of leucine enkephalin-alka(e)nal-glycerophosphatidylcholine adducts by tandem mass spectrometry

J Am Soc Mass Spectrom. 2006 May;17(5):657-60. doi: 10.1016/j.jasms.2006.01.006. Epub 2006 Mar 6.

Abstract

The covalent interactions between peptides and lipid oxidation products, with formation of Schiff and Michael adducts, are known to occur during free radical oxidative damage. In this study, leucine-enkephalin-glycerophosphatidylcholine alka(e)nal adducts were analyzed by electrospray tandem mass spectrometry (MS/MS). Upon collision-induced dissociation of the Leucine enkephalin-2-(9-oxo-nonanoyl)-1-palmitoyl-3-glycerophosphatidylcholine, an alkanal Schiff adduct observed at m/z 1187.7, the main product ions were attributed to the phosphocholine polar head and loss of the peptide. Also, product ions resulting from characteristic losses of phosphatidylcholines and cleavages of the peptide chain (mainly b-type) were observed. Additional product ions formed by combined peptide and phosphatidylcholine fragmentations were identified. The fragmentation pattern of the leucine enkephalin-alkanal Schiff adduct and the leucine enkephalin-alkenal phosphatidylcholine Schiff and Michael adducts were similar, although the loss of the peptide for the Michael adduct should occur through a distinct mechanism. These fragmentation pathways differ greatly from those described for peptide-lipid Schiff and Michael adducts, in which only peptide chain cleavages are reported, probably due to charge retention in the glycerophosphatidylcholine polar head in peptide-glycerophosphatidylcholine adducts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross-Linking Reagents / chemistry*
  • Enkephalin, Leucine / chemistry*
  • Neurotransmitter Agents / chemistry*
  • Peptides / chemistry*
  • Phosphatidylcholines / chemistry
  • Spectrometry, Mass, Electrospray Ionization
  • Tandem Mass Spectrometry / methods*

Substances

  • Cross-Linking Reagents
  • Neurotransmitter Agents
  • Peptides
  • Phosphatidylcholines
  • Enkephalin, Leucine
  • 1-palmitoyl-2-linoleoylphosphatidylcholine