UNIT 17.14A Preparation of Glycopeptides

  1. Leland D. Powell

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1714as32

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Powell, L. D. 2001. Preparation of Glycopeptides. Current Protocols in Molecular Biology. 32:III:17.14A:17.14.1–17.14.9.

Author Information

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

Publication History

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


Generation of glycopeptides from glycoproteins is frequently useful when analyzing a protein's oligosaccharide side chains. Freed from the bulk of the polypeptide backbone by proteolysis, glycopeptides can be characterized by a variety of techniques. Extensive proteolysis with pronase or proteinase K results in oligosaccharides with one or a few amino acid residues attached. This technique, detailed in this unit, is often employed as a first step in characterizing oligosaccharides on very large glycoproteins such as proteoglycans and mucins. Limited proteolysis with a specific endoproteinase (e.g., trypsin, a-chymotrypsin, and V8 protease) is also described, and leaves a larger peptide attached to the oligosaccharide. The resulting glycopeptides are generally suitable substrates for Peptide:N-glycosidase F, an enzyme useful in defining oligosaccharide-peptide linkages. Additionally, they can be separated by reversed-phase chromatography, resulting in a glycopeptide map that is analogous to a peptide map, and used for detection of glycosylation sites.