Neuraminidase Sp

Sialidase, N-acetylneuraminate glycohydrolase, Exo-alpha-sialidase

α(2-3) Neuraminidase Sp cleaves exclusively the non-reducing terminal α(2-3) unbranched sialic acid residues from complex carbohydrates and glycoproteins. There is no detectable activity on α(2-6) or α(2-8) linkages or on branched α(2-3) linkages . To cleave all non-reducing terminal sialic acid residues including branched sialic acids (linked to an internal residue) from complex carbohydrates and glycoproteins, use α(2-3,6,8,9) Neuraminidase Au (E-S001).

Source recombinant Streptococcus pneumoniae in E. Coli

EC 3.5.1.18

Contents
Neuraminidase Sp in 50 mM sodium phosphate, pH 7.5

Included with 20 µL and 60 µL pack sizes:
5x Reaction Buffer 250 mM sodium phosphate, pH 6.0

Specific Activity ≥ 150 U/mg
Activity ≥ 5 U/ml

Molecular weight~75,000 daltons

pH optimum 6.0

Suggested usage 1. Add up to 100 μg of glycoprotein or 1 nmol of oligosaccharide to tube.
2. Add water to 14 μL
3. Add 4 μL 5X Reaction Buffer.
4. Add 2 μL α (2-3) Neuraminidase.
5. Incubate at 37°C for 1 hour

Desialylation may be monitored by SDS-PAGE if the size differential between native and desialylated protein is sufficient for detection.

Specifictity Cleaves the non-reducing terminal alpha-(2-3) unbranched sialic acid residues from complex carbohydrates and glycoproteins.

Specific Activity Defined as the amount of enzyme required to produce 1 µmole of methylumbelliferone in 1 minute at 37˚C, pH 5.0 from MU-NANA [2′-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid].

Storage Store enzyme at 4˚C.

Stability Stable at least 24 months when stored properly. Several days exposure to ambient tempertures will not reduce activity.

Neuraminidase Sp References

Corfield, A. P., H. Higa, J. C. Paulson and R. Schauer. The specificity of viral and bacterial sialidases for alpha(2-3) and alpha(2-6)-linked sialic acids in glycoproteins. Biochim Biophys Acta 744: 121-12 6 (1983).

Dwek, R. A., C. J. Edge, D. J. Harvey, M. R. Wormald and R. B. Parekh. Analysis of glycoprotein-associated oligosaccharides. Ann Rev Biochem 62: 65-100 (1993).

Kobata, A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem 100: 1-14 (1979).

Glasgow, L R., J. C . Paulson and R. L. Hill. Systematic purification of five glycosidases from Streptococcus pneumoniae. J. Biol Chem 252: 8615-8623 (1977).

Kelly, R. T., D. Greiff and S. Farmer. Neuraminidase activity in Diplococcus pneumoniae. J Bacteriol 91: 601-3 (1965).

Prime, S., J. Dearnley , A. M. Venton, R. B. Parekh and C. J. Edge. Oligosaccharide sequencing based on exo- and endoglycosidase digestion and liquid chromatographic analysis of the products. J Chromatogr A 720: 263-274 (1996).