High-sensitivity electron capture dissociation tandem FTIRC mass spectrometry of microelectrosprayed peptides

K. Håkansson, Mark Emmett, C. L. Hendrickson, A. G. Marshall

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Electron capture dissociation (ECD) has previously been shown by other research groups to result in greater peptide sequence coverage than other ion dissociation techniques and to localize labile posttranslational modifications. Here, ECD has been achieved for 10-13-mer peptides microelectrosprayed from 10 nM (10 fmol/μL) solutions and for tryptic peptides from a 50 nM unfractionated digest of a 28-kDa protein. Tandem Fourier transform ion cyclotron resonance (FTICR) mass spectra contain fragment ions corresponding to cleavages at all possible peptide backbone amine bonds, except on the N-terminal side of proline, for substance P and neurotensin. For luteinizing hormone-releasing hormone, all but two expected backbone amine bond cleavages are observed. The tandem FTICR mass spectra of the tryptic peptides contain fragment ions corresponding to cleavages at 6 of 12 (1545.7-Da peptide) and 8 of 21 (2944.5-Da peptide) expected backbone amine bonds. The present sensitivity is 200-2000 times higher than previously reported. These results show promise for ECD as a tool to produce sequence tags for identification of peptides in complex mixtures available only in limited amounts, as in proteomics.

Original languageEnglish (US)
Pages (from-to)3605-3610
Number of pages6
JournalAnalytical Chemistry
Volume73
Issue number15
DOIs
StatePublished - Aug 1 2001
Externally publishedYes

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Mass spectrometry
Peptides
Electrons
Ions
Amines
Cyclotron resonance
Fourier transforms
Neurotensin
Substance P
Complex Mixtures
Proline
Gonadotropin-Releasing Hormone
Proteins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

High-sensitivity electron capture dissociation tandem FTIRC mass spectrometry of microelectrosprayed peptides. / Håkansson, K.; Emmett, Mark; Hendrickson, C. L.; Marshall, A. G.

In: Analytical Chemistry, Vol. 73, No. 15, 01.08.2001, p. 3605-3610.

Research output: Contribution to journalArticle

Håkansson, K. ; Emmett, Mark ; Hendrickson, C. L. ; Marshall, A. G. / High-sensitivity electron capture dissociation tandem FTIRC mass spectrometry of microelectrosprayed peptides. In: Analytical Chemistry. 2001 ; Vol. 73, No. 15. pp. 3605-3610.
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