UNIT 17.13A Endoglycosidase and Glycoamidase Release of N-Linked Oligosaccharides
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Freeze, H. H. 2001. Endoglycosidase and Glycoamidase Release of N-Linked Oligosaccharides. Current Protocols in Molecular Biology. 48:17.13A.1–17.13A.26.
- Published Online: 1 MAY 2001
- Published Print: OCT 1999
This is not the most recent version of the article. View current version (1 JAN 2010)
Carbohydrate chain modifications are often used to monitor glycoprotein movement through the secretory pathway. This is because stepwise sugar-chain processing is unidirectional and generally corresponds to the forward or anterograde movement of proteins. This unit offers a group of techniques that will help analyze the general structure of carbohydrate chains on a protein and, therefore, oligosaccharide processing mileposts. The sugar chains themselves are not analyzed, but their presence and structure are inferred from gel mobility differences after one or more enzymatic digestions. This approach is most often used in combination with [35S]Met pulse-chase metabolic labeling protocols, but they can be applied to any suitably labeled protein (e.g., biotinylated or 125I-labeled).