Automated N-glycopeptide identification using a combination of single- and tandem-MS

J Proteome Res. 2007 Oct;6(10):3995-4005. doi: 10.1021/pr070239f. Epub 2007 Aug 30.

Abstract

We describe Peptoonist, a program that can automatically identify the glycans (sugars) present at each N-glycosylation site of a protein. The input to Peptoonist is a series of mass spectra, both MS and MS/MS, obtained from a liquid chromatography (LC) run of proteolytically digested purified glycoproteins. The program uses MS/MS to identify glycosylated peptides and single-MS to identify the N-glycans present on each of these peptides, at least to the level of monosaccharide composition. We validate the program on an LC run of mouse zona pellucida proteins that had been intensively hand annotated by a human expert. Our program doubled the number of glycopeptide identifications, and also found several possible errors in the hand annotation. In addition, it automatically made most of the same glycan isomer identifications as the expert annotator.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoanalysis
  • Egg Proteins / analysis
  • Glycopeptides / analysis*
  • Glycosylation
  • Humans
  • Isomerism
  • Mass Spectrometry / methods*
  • Membrane Glycoproteins / analysis
  • Mice
  • Molecular Sequence Data
  • Receptors, Cell Surface / analysis
  • Software
  • Tandem Mass Spectrometry
  • Zona Pellucida Glycoproteins

Substances

  • Egg Proteins
  • Glycopeptides
  • Membrane Glycoproteins
  • Receptors, Cell Surface
  • Zona Pellucida Glycoproteins