Endoglycosidase H

Endoglycosidase H (Endo H) is a recombinant glycosidase cloned from Streptomyces plicatus and overexpressed in E. coli. Endo H cleaves the chitobiose core of high mannose and a limited number of hybrid oligosaccharides from N-linked glycoproteins. It does not cleave complex glycans. Enzymatic cleavage is between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, leaving one N-acetylglucosamine residue on the asparagine. This is in contrast to PNGase F, which cleaves all asparagine-linked oligosaccharides.

Unit Definition: One unit is defined as the amount of enzyme required to remove >95% of the carbohydrate from 10μg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10μl.

• Determining the state of protein glycosylation
• Characterizing glycan structure

1. Maley, F. et al. (1989) Anal. Biochem. 180, 195–204.

References using Endo H to monitor protein trafficking:

1. Taner, S. et al. (2011) J. Immunol. 186, 62–72.
2. Beyer, S. et al. (2011) Endo. 152, 782–92.
3. Chen, Z. et al. (2011) J. Exper. Med. 208, 2633–40.
4. Iavarone, C. et al. (2011) J. Immunol. 186, 4213–22.
5. Guo, Y. et al. (2011) J. Exper. Med. 208, 2083–98.