UNIT 17.14B Detection of Individual Glycosylation Sites on Glycoproteins

  1. Leland D. Powell

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1714bs32

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Powell, L. D. 2001. Detection of Individual Glycosylation Sites on Glycoproteins. Current Protocols in Molecular Biology. 32:III:17.14B:17.14.10–17.14.15.

Author Information

  1. University of California San Diego, School of Medicine, La Jolla, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1995


In this unit, glycopeptides generated by endopeptidase digestion are first separated by reversed-phase chromatography. The presence of hydrophilic, negatively charged oligosaccharides shortens retention times, causing glycopeptides to elute in considerably broader peaks than do peptides, so by following the elution profile either radiochemically or colorimetrically, the peaks corresponding to unique glycopeptides can be identified. With proper controls, the number of peaks will correspond to the number of different glycosylation sites. The eluted fractions are suitable for analysis by lectin chromatography, and the peptide sugar linkage can be defined either by endoglycosidase digestion or chemical cleavage. Oligosaccharides freed from the peptide according to the methods described in this unit can be characterized by size or charge, techniques not generally applicable with glycopeptides.