On-Line Liquid Chromatography Electron Capture Dissociation for the Characterization of Phosphorylation Sites in Proteins

  • Steve M.M. M. Sweet
  • Helen J. Cooper
Part of the Methods in Molecular Biology™ book series (MIMB, volume 527)


Electron capture dissociation (ECD) allows fragmentation of the phosphopeptide backbone while keeping the labile phospho-amino acid intact. This feature of ECD fragmentation, coupled with the acquisition of mass spectra at high mass accuracy, makes ECD well-suited to phosphorylation mapping. The following methods are designed to focus ECD events on phosphopeptides within a complex peptide sample, either by using phosphoric acid neutral loss peaks as a trigger or by targeted analysis of predetermined precursor masses.

Key words

ECD Neutral loss Phosphorylation FT-ICR LTQ-FT Xcalibur Site localization AGC Targeted OMSSA 



We are grateful to Lewis Geer for assistance with the OMSSA algorithm for ECD. This work was funded by the EU (FP6 Endotrack; (SMMS) and the Wellcome Trust (074131) (HJC).


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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Steve M.M. M. Sweet
    • 1
  • Helen J. Cooper
    • 2
  1. 1.University of Birmingham, School of BiosciencesBirmingham
  2. 2.School of Biosciences, University of BirminghamBirmingham

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