Global proteomic profiling of phosphopeptides using electron transfer dissociation tandem mass spectrometry

Henrik Molina, David M. Horn, Ning Tang, Suresh Mathivanan, Akhilesh Pandey

Research output: Contribution to journalArticle

Abstract

Electron transfer dissociation (ETD) is a recently introduced mass spectrometric technique that provides a more comprehensive coverage of peptide sequences and posttranslational modifications. Here, we evaluated the use of ETD for a global phosphoproteome analysis. In all, we identified a total of 1,435 phosphorylation sites from human embryonic kidney 293T cells, of which 1,141 (≈80%) were not previously described. A detailed comparison of ETD and collision-induced dissociation (CID) modes showed that ETD identified 60% more phosphopeptides than CID, with an average of 40% more fragment ions that facilitated localization of phosphorylation sites. Although our data indicate that ETD is superior to CID for phosphorylation analysis, the two methods can be effectively combined in alternating ETD and CID modes for a more comprehensive analysis. Combining ETD and CID, from this single study, we were able to identify 80% of the known phosphorylation sites in > 1,000 phosphorylated peptides analyzed. A hierarchical clustering of the identified phosphorylation sites allowed us to discover 15 phosphorylation motifs that have not been reported previously. Overall, ETD is an excellent method for localization of phosphorylation sites and should be an integral component of any strategy for comprehensive phosphorylation analysis.

Original languageEnglish (US)
Pages (from-to)2199-2204
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume104
Issue number7
DOIs
StatePublished - Feb 13 2007

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Phosphopeptides
Tandem Mass Spectrometry
Proteomics
Phosphorylation
Electrons
Peptides
HEK293 Cells
Post Translational Protein Processing
Cluster Analysis
Ions
Kidney

Keywords

  • Bioinformatics
  • Motifs
  • Phosphorylation
  • Signal transduction
  • Systems biology

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Global proteomic profiling of phosphopeptides using electron transfer dissociation tandem mass spectrometry. / Molina, Henrik; Horn, David M.; Tang, Ning; Mathivanan, Suresh; Pandey, Akhilesh.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 7, 13.02.2007, p. 2199-2204.

Research output: Contribution to journalArticle

Molina, Henrik ; Horn, David M. ; Tang, Ning ; Mathivanan, Suresh ; Pandey, Akhilesh. / Global proteomic profiling of phosphopeptides using electron transfer dissociation tandem mass spectrometry. In: Proceedings of the National Academy of Sciences of the United States of America. 2007 ; Vol. 104, No. 7. pp. 2199-2204.
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