LC/MSn for glycoprotein analysis: N-linked glycosylation analysis and peptide sequencing of glycopeptides

Methods Mol Biol. 2009:534:239-48. doi: 10.1007/978-1-59745-022-5_18.

Abstract

Liquid chromatography/multiple-stage mass spectrometry (LC/MS( n )) is an effective means for the site-specific glycosylation analysis of a limited quantity of glycoproteins, such as gel-separated proteins. Generally, a tryptic digest of the glycoprotein is separated by reversed-phase LC, and peptide sequencing and glycosylation analysis are achieved with on-line MS( n ). In this chapter, a protocol for the LC/MS/MS/MS of a proteolytic digest of a gel-separated glycoprotein is described.

Publication types

  • Review

MeSH terms

  • Carbohydrate Sequence
  • Chromatography, Liquid / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Glycoproteins / analysis*
  • Glycoproteins / chemistry
  • Glycoproteins / metabolism
  • Glycosylation*
  • Humans
  • Methylation
  • Molecular Sequence Data
  • Protein Processing, Post-Translational / physiology
  • Reducing Agents / pharmacology
  • Sequence Analysis, Protein / methods*
  • Tandem Mass Spectrometry / methods*

Substances

  • Glycoproteins
  • Reducing Agents