TitleExploring ECD on a Benchtop Q Exactive Orbitrap Mass Spectrometer.
Publication TypeJournal Article
Year of Publication2018
AuthorsFort KL, Cramer CN, Voinov VG, Vasil'ev YV, Lopez NI, Beckman JS, Heck AJR
JournalJ Proteome Res
Volume17
Issue2
Pagination926-933
Date Published2018 02 02
ISSN1535-3907
KeywordsAmino Acid Sequence, Disulfides, Electrons, Equipment Design, Humans, Ions, Myoglobin, Peptide Fragments, Protein Processing, Post-Translational, Proteolysis, Proteomics, Substance P, Tandem Mass Spectrometry, Ubiquitin
Abstract

As the application of mass spectrometry intensifies in scope and diversity, the need for advanced instrumentation addressing a wide variety of analytical needs also increases. To this end, many modern, top-end mass spectrometers are designed or modified to include a wider range of fragmentation technologies, for example, ECD, ETD, EThcD, and UVPD. Still, the majority of instrument platforms are limited to more conventional methods, such as CID and HCD. While these latter methods have performed well, the less conventional fragmentation methods have been shown to lead to increased information in many applications including middle-down proteomics, top-down proteomics, glycoproteomics, and disulfide bond mapping. We describe the modification of the popular Q Exactive Orbitrap mass spectrometer to extend its fragmentation capabilities to include ECD. We show that this modification allows ≥85% matched ion intensity to originate from ECD fragment ion types as well as provides high sequence coverage (≥60%) of intact proteins and high fragment identification rates with ∼70% of ion signals matched. Finally, the ECD implementation promotes selective disulfide bond dissociation, facilitating the identification of disulfide-linked peptide conjugates. Collectively, this modification extends the capabilities of the Q Exactive Orbitrap mass spectrometer to a range of new applications.

DOI10.1021/acs.jproteome.7b00622
Alternate JournalJ. Proteome Res.
PubMed ID29249155
PubMed Central IDPMC5799867
Grant ListR44 GM122131 / GM / NIGMS NIH HHS / United States