Table 2 Summary of glycosidases and running conditions used for digestion of mosquito cell line glycoproteins.
|
Enzyme (enzyme number) | Abbreviation | Cleavage site |
Enzyme Conc. | Incubation Buffer (pH) | Reference |
|---|---|---|---|---|---|
|
Peptide-
N
-glycosidase F
(EC 3.2.2.18) | PNGase F |
N-linked oligosaccharides-Asn | 50 U | 0.5 M sodium phosphate (7.5) | [59] |
|
Peptide-
N
-glycosidase A
(EC 3.5.1.52) | PNGase A |
N-linked oligosaccharides-Asn | 2.5 mU | 50 mM sodium citrate, sodium phosphate (5.0) | [60] |
|
Endo-β-glucosaminidase H
(EC 3.2.1.96) | Endo H |
Oligomannose/ hybrid oligosaccharides-Asn | 50 U | 0.5 M sodium citrate (5.5) | [61] |
|
Endo-α-acetylgalactosaminidase
(EC 3.2.1.92/3.2.1.110) | O-glycanase™ | Galβ1-3GalNAcα1-Ser/Thr | 82-112 mU | 10 mM sodium citrate, sodium phosphate (6.0) | [62] |
|
α-
N
-acetylgalactosaminidase
(EC 3.2.1.49) | α-AGA | GalNAcα1-3R/Ser/Thr | 625 mU | 100 mM sodium citrate, sodium phosphate (3.5-4) | [63] |
|
β-Galactosidase
(EC 3.2.1.23) | - |
Galβ1-3GlcNAc -Glc/Man | 4 mU | 20 mM sodium citrate, sodium phosphate (6.0) | [64] |
|
Neuraminidase
(EC 3.2.1.18) | - | Sialic acidα2-3,6,8R | 25 mU | 0.5 M sodium citrate (4.5) | [65] |