A strategy for identification of oligosaccharide structures using observational multistage mass spectral library

Anal Chem. 2005 Aug 1;77(15):4719-25. doi: 10.1021/ac048350h.

Abstract

Glycosylation is the most widespread posttranslational modification in eukaryotes; however, the role of oligosaccharides attached to proteins has been little studied because of the lack of a sensitive and easy analytical method for oligosaccharide structures. Recently, tandem mass spectrometric techniques have been revealing that oligosaccharides might have characteristic signal intensity profiles. We describe here a strategy for the rapid and accurate identification of the oligosaccharide structures on glycoproteins using only mass spectrometry. It is based on a comparison of the signal intensity profiles of multistage tandem mass (MSn) spectra between the analyte and a library of observational mass spectra acquired from structurally defined oligosaccharides prepared using glycosyltransferases. To smartly identify the oligosaccharides released from biological materials, a computer suggests which ion among the fragment ions in the MS/MS spectrum should yield the most informative MS3 spectrum to distinguish similar oligosaccharides. Using this strategy, we were able to identify the structure of N-linked oligosaccharides in immunoglobulin G as an example.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Databases, Factual*
  • Glycoproteins / analysis
  • Glycoproteins / chemistry
  • Glycoproteins / metabolism
  • Humans
  • Immunoglobulin G / chemistry
  • Mass Spectrometry / methods*
  • Molecular Conformation
  • Molecular Sequence Data
  • Oligosaccharides / analysis*
  • Oligosaccharides / chemistry*

Substances

  • Glycoproteins
  • Immunoglobulin G
  • Oligosaccharides