Enrichment of glycopeptides for glycan structure and attachment site identification

Nat Methods. 2009 Nov;6(11):809-11. doi: 10.1038/nmeth.1392. Epub 2009 Oct 18.


We present a method to enrich for glycoproteins from proteomic samples. Sialylated glycoproteins were selectively periodate-oxidized, captured on hydrazide beads, trypsinized and released by acid hydrolysis of sialic acid glycosidic bonds. Mass spectrometric fragment analysis allowed identification of glycan structures, and additional fragmentation of deglycosylated ions yielded peptide sequence information, which allowed glycan attachment site and protein identification. We identified 36 N-linked and 44 O-linked glycosylation sites on glycoproteins from human cerebrospinal fluid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cerebrospinal Fluid Proteins / analysis*
  • Glycopeptides / analysis
  • Glycoproteins / chemistry*
  • Glycoproteins / isolation & purification*
  • Glycosylation
  • Humans
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / metabolism
  • Polysaccharides / analysis
  • Polysaccharides / chemistry
  • Proteomics
  • Tandem Mass Spectrometry


  • Cerebrospinal Fluid Proteins
  • Glycopeptides
  • Glycoproteins
  • Polysaccharides
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase