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Sites of glycosylation

N- and O-linked site mapping, including 18O-labelling and LC-MS isotopic shift monitoring.

  • Determination of N– and O-linked site mapping is often a crucial part of glycoprotein characterization.
  • Enzymatic deglycosylation in the presence of 18O-labeled water results in an 18O label on the site of the recently removed N-glycan. For O-linked glycosylation, BEMAD (β- elimination followed by Michael addition with dithiothreitol) may be used.
  • Isotopic shift monitoring using LC-MS methodology helps provide information regarding the position and extent of N-linked glycosylation.
Diagram shows peptide fragmentation methods (CID, HCD, ETD) with resulting ion types (b, y, c, z) identified after each process. Peptide sequence and fragmentations are illustrated.

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