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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)

Summary

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 

Notes

Acknowledgments

We are grateful to Lewis Geer for assistance with the OMSSA algorithm for ECD. This work was funded by the EU (FP6 Endotrack; www.endotrack.org) (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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