Comprehensive comparison of collision induced dissociation and electron transfer dissociation

Henrik Molina, Rune Matthiesen, Kumaran Kandasamy, Akhilesh Pandey

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Electron transfer dissociation (ETD) is a recently introduced mass spectrometric technique which has proven to be an excellent tool for the elucidation of labile posttranslational modifications such as phosphorylation and O-GlcNAcylation of serine and threonine residues. However, unlike collision induced dissociation (CID), which has been studied for decades, the intricacies of ETD-based fragmentation have not yet been firmly established or systematically addressed. In this analysis, we have systematically compared the CID and ETD fragmentation patterns for the large majority of the peptides that do not contain such labile modifications. Using a standard 48 protein mix, we were able to measure false-positive rates for the experiments and also assess a large number of peptides for a detailed comparison of CID and ETD fragmentation pattern. Analysis of ∼19 000 peptides derived from both standard proteins and complex protein samples revealed that (i) CID identified 50% more peptides than ETD; (ii) ETD resulted in ∼20% increase in amino acid sequence coverage over CID; and (iii) combining CID and ETD fragmentation increased the sequence coverage for an average tryptic peptide to 92%. Interestingly, our analysis revealed that nearly 60% of all ETD-identified peptides carried two positive charges, which is in sharp contrast to what has been generally accepted. We also present a novel strategy for automatic validation of peptide assignments based on identification of a peptide by consecutive CID and ETD fragmentation in an alternating mode.

Original languageEnglish (US)
Pages (from-to)4825-4835
Number of pages11
JournalAnalytical Chemistry
Volume80
Issue number13
DOIs
StatePublished - Jul 1 2008
Externally publishedYes

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Electrons
Peptides
Phosphorylation
Proteins
Threonine
Serine
Amino Acids

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Comprehensive comparison of collision induced dissociation and electron transfer dissociation. / Molina, Henrik; Matthiesen, Rune; Kandasamy, Kumaran; Pandey, Akhilesh.

In: Analytical Chemistry, Vol. 80, No. 13, 01.07.2008, p. 4825-4835.

Research output: Contribution to journalArticle

Molina, Henrik ; Matthiesen, Rune ; Kandasamy, Kumaran ; Pandey, Akhilesh. / Comprehensive comparison of collision induced dissociation and electron transfer dissociation. In: Analytical Chemistry. 2008 ; Vol. 80, No. 13. pp. 4825-4835.
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