Table of enzymes for glycan analysis
| EC number | Specificity (cleavage target) | Analytical uses | |
|---|---|---|---|
| Endoglycosidases | |||
| Endoglycosidase H (Endo H) | 3.2.1.96 | between the two GlcNAc residues in the core of oligomannose or hybrid N-glycans | detection of N-glycosylation; release of glycans |
| Endoglycosidase F2 (Endo F) | 3.2.1.96 | between the two GlcNAc residues in the core of oligomannose or biantennary N-glycans | detection of N-glycosylation; release of glycans |
| Peptide-N-glycosidase F (PNGase F), or glycopeptidase F | 3.5.1.52 | between Asn and oligomannose, complex, or hybrid N-glycans; requires at least one amino acid at both the amino terminal and carboxy terminal of Asn | may not work if core is α1-3-fucosylated |
| Peptide-N-glycosidase A (PNGase A), or glycopeptidase A | 3.5.1.52 | between Asn and oligomannose, complex, or hybrid N-glycans; requires at least one amino acid at both the amino terminal and carboxy terminal of Asn | will work if core is α1-3-fucosylated; efficacy for use on mammalian glycoproteins poorly defined |
| Endo-β-galactosidases | 3.2.1.102 | between Gal and GlcNAc in poly-N-acetyllactosmines | detection of poly-N-acetyllactosamines and some keratan sulfates |
| Endoneuraminidases (endo-α-sialidase) | 3.2.1.129 | between Sia units of polysialic acids | detection of polysialic acids |
| Endo-α-N-acetylgalactosaminidase (O-glycanase) | 3.2.1.97 | Galβ-GalNAcα- from serine or threonine | detects O-glycosidation. May need prior treatment with other enzymes to expose the disaccharide core |
| Exoglycosidases | |||
| Sialidases, neuraminidases | 3.2.1.18 | nonreducing terminal Sia | |
| Fucosidase | 3.2.1.51 | nonreducing terminal Fuc | some types and linkages can be resistant |
| α1-2 Fucosidase | 3.2.1.63 | nonreducing terminal Fuc-linked α1-2 | |
| β-Galactosidase | 3.2.1.23 | nonreducing terminal β-Gal | |
| α-Mannosidase | 3.2.1.24 | nonreducing terminal α-Man | |
| Glycosaminoglycan Lyases | |||
| Chondroitinase ABC | 4.2.2.4 | chondroitin sulfates A and C and dermatan sulfate to disaccharides | detection and characterization of chondroitins, purification of other glycosaminoglycans |
| Chondroitinase AC | 4.2.2.5 | chondroitin sulfates A and C to disaccharides | detection and characterization of chondroitins A and C |
| Chondroitinase B | 4.2.2.19 | dermatan sulfate (chondroitin sulfate B) to disaccharides | detection and characterization of dermatan sulfate |
| Heparinase, heparin lyase, Heparinase I | 4.2.2.7 | between N-sulfated glucosamine and 2-O-sulfated iduronate residues. | detection and characterization of heparin and heparan sulfate |
| Heparin sulfate lyase, heparinase III (also called Heparitinase I), Heparinase II | 4.2.2.8 | between GlcNAc and GlcA residues. Heparinase I, II, and III often used in combination. | detection and characterization of heparin and heparan sulfate |
| Keratanase | 3.2.1.103 | between Gal and GlcA | detection and characterization of keratan sulfate |
| Keratanase II | 3.2.1.103 | between 6-O-sulfated GlcNAc and Gal (±6-O-sulfate) | detection and characterization of keratan sulfate |
EC, Enzyme Commission.
From: Chapter 50, Structural Analysis of Glycans
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