Journal of the American Society for Mass Spectrometry

, Volume 17, Issue 10, pp 1429–1436

The effect of radical trap moieties on electron capture dissociation spectra of substance P


  • Marina A. Belyayev
    • Department of ChemistryBoston University
  • Jason J. Cournoyer
    • Department of ChemistryBoston University
  • Cheng Lin
    • Department of ChemistryBoston University
    • Department of ChemistryBoston University

DOI: 10.1016/j.jasms.2006.06.008

Cite this article as:
Belyayev, M.A., Cournoyer, J.J., Lin, C. et al. J Am Soc Mass Spectrom (2006) 17: 1429. doi:10.1016/j.jasms.2006.06.008


To further test the hypothesis that electron capture dissociation (ECD) involves long-lived radical intermediates and radical migration occurs within these intermediates before fragmentation, radical trap moieties were attached to peptides with the assumption that they would reduce fragmentation by decreasing the mobility of the radical. Coumarin labels were chosen for the radical traps, and unlabeled, singly-labeled, and doubly-labeled Substance P were analyzed by ECD. The results demonstrated a correlation between the number and position of tags on the peptide and the intensity of side-chain cleavages observed, as well as an inverse correlation between the number of tags on the peptide and the intensity of backbone cleavages. Addition of radical traps to the peptide inhibits backbone cleavages, suggesting that either radical mobility is required for these cleavages, or new noncovalent interactions prevent separation of backbone cleavage fragments. The enhancement of side-chain cleavages and the observation of new side-chain cleavages associated with aromatic groups suggest that the gas-phase conformation of this peptide is substantially distorted from untagged Substance P and involves previously unobserved interactions between the coumarin tags and the phenylalanine residues. Furthermore, the use of a double resonance (DR)-ECD experiment showed that these side-chain losses are all products of long-lived radical intermediate species, which suggests that steric hindrance prevents the coumarin-localized radical from interacting with the backbone while simultaneously increasing the radical rearrangements with the side chains.

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© American Society for Mass Spectrometry 2006