Obtaining complementary polypeptide sequence information from a single precursor ion packet via sequential ion mobility-resolved electron transfer and vibrational activation

Deepali Rathore, Forouzan Aboufazeli, Eric D. Dodds

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Tandem mass spectrometry (MS/MS) is now well-known as a powerful tool for characterizing the primary structures of peptides and proteins; however, in many cases the use of but a single dissociation method provides only a partial view of the amino acid sequences and post-translational modification patterns of polypeptides. While the application of multiple fragmentation methods can be more informative, this introduces the burden of acquiring multiple MS/MS spectra per analyte, thus reducing the effective duty cycle of such methods. In this work, initial proof-of-concept is provided for a method designed to overcome these barriers. This method relies on the complementary fragmentation information that can be provided by performing collision-induced dissociation (CID) and electron transfer dissociation (ETD) in concert, while also taking advantage of an ion mobility (IM) dimension to temporally resolve the occurrence of CID and ETD when applied to a single accumulated packet of precursor ions. In this way, the significant proportion of the precursor ion population that remains unreacted in ETD experiments is subjected to CID rather than being fruitlessly discarded. In addition, the two distinct fragmentation spectra can be extracted from their corresponding IM domains to render readily interpretable individual fragmentation spectra. This scheme was demonstrated for several polypeptides ranging from 1.3 to 8.6 kDa in molecular weight. In each case, IM-resolved CID and ETD events resulted in b/y and c/z ions, respectively, which each covered both unique and overlapping sequence information. These findings demonstrate that the combination of CID and ETD can be achieved with greater utilization of the available ion population and little or no loss of duty cycle.

Original languageEnglish (US)
Pages (from-to)7175-7183
Number of pages9
JournalAnalyst
Volume140
Issue number21
DOIs
StatePublished - Nov 7 2015

Fingerprint

Electron mobility
Polypeptides
Chemical activation
Electrons
Ions
electron
Peptides
ion
collision
fragmentation
Amino Acid Sequence
Post Translational Protein Processing
Tandem Mass Spectrometry
peptide
Population
amino acid
mass spectrometry
Molecular Weight
method
protein

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

Obtaining complementary polypeptide sequence information from a single precursor ion packet via sequential ion mobility-resolved electron transfer and vibrational activation. / Rathore, Deepali; Aboufazeli, Forouzan; Dodds, Eric D.

In: Analyst, Vol. 140, No. 21, 07.11.2015, p. 7175-7183.

Research output: Contribution to journalArticle

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