Sequencing-Grade De novo Analysis of MS/MS Triplets (CID/HCD/ETD) From Overlapping Peptides

peer-reviewed · Journal of Proteome Research · 2013

peer-reviewed · Journal of Proteome Research · 2013. Adrian Guthals et al. Full-length de novo sequencing of unknown proteins remains a challenging open problem. Traditional methods…
Date 2013-06-07
Type peer-reviewed
Venue Journal of Proteome Research
Publisher American Chemical Society (ACS)
Contribution algorithm
DOI 10.1021/pr400173d
Citations (OpenAlex) 73
Venue 2-year citedness 3.48

Abstract

Full-length de novo sequencing of unknown proteins remains a challenging open problem. Traditional methods that sequence spectra individually are limited by short peptide length, incomplete peptide fragmentation, and ambiguous de novo interpretations. We address these issues by determining consensus sequences for assembled tandem mass (MS/MS) spectra from overlapping peptides (e.g., by using multiple enzymatic digests). We have combined electron-transfer dissociation (ETD) with collision-induced dissociation (CID) and higher-energy collision-induced dissociation (HCD) fragmentation methods to boost interpretation of long, highly charged peptides and take advantage of corroborating b/y/c/z ions in CID/HCD/ETD. Using these strategies, we show that triplet CID/HCD/ETD MS/MS spectra from overlapping peptides yield de novo sequences of average length 70 AA and as long as 200 AA at up to 99% sequencing accuracy.

Authors

  1. Adrian Guthals · Mapp Biopharmaceutical, Inc., University of California San Diego
  2. Karl R. Clauser · Broad Institute of the Massachusetts Institute of Technology and Harvard, Millennium Pharmaceuticals
  3. Ari Frank · Affectivon, Inc., Max Planck Institute of Molecular Cell Biology and Genetics, University of California San Diego
  4. Nuno Bandeira · University of California San Diego

Methods and tools

  • MS/MS Triplets: Sequencing-grade de novo analysis from overlapping CID/HCD/ETD MS/MS triplets.

Cites (11)

Cited by (13)

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