Rapid de-O-glycosylation concomitant with peptide labeling using microwave radiation and an alkyl amine base.

Academic Article

Abstract

  • Procedures are detailed for a quantitative release of O-linked glycans from peptides that now provide a shorter reaction time, a possible identification of O-linked sites, and a quantification of all reaction products. The release was initiated by a mild base, dimethylamine, and accelerated by microwave radiation. Differential analysis using standard glycoproteins has shown improved release efficiency concurrent with facile incorporation of dimethylamine into the former O-linked sites. In situ glycan reduction insures protection against peeling and is synchronous with subsequent studies by high performance MS(n) sequencing. The protocols were established with a synthetic O-GlcNAc peptide that would mimic the linkage chemistry and applied to a well characterized glycoprotein bovine fetuin with both N- and O-linked glycans and a highly glycosylated swine mucin.
  • Authors

  • Maniatis, Stephanie
  • Zhou, Hui
  • Reinhold, Vernon
  • Status

    Publication Date

  • March 15, 2010
  • Keywords

  • Amino Acid Sequence
  • Animals
  • Cattle
  • Dimethylamines
  • Glycoproteins
  • Glycosylation
  • Microwaves
  • Peptides
  • Polysaccharides
  • Swine
  • Digital Object Identifier (doi)

    Start Page

  • 2421
  • End Page

  • 2425
  • Volume

  • 82
  • Issue

  • 6