Chemical degradation ~Smith degradation
Smith degradation, which consists of partial periodate oxidation and aldehyde-reduction, was developed by Smith et al. for linkage analysis of carbohydrates. 1, 2) The periodate oxidation cleaves between diol. The method was improved to high sensitivity with fluorescence-tagged carbohydrates by Omichi and Hase. 3)
sodium metaperiodate (Wako Pure Chemicals, Osaka, Japan)
sodium acetate (Wako Pure Chemicals, Osaka, Japan)
sodium borohydride (Wako Pure Chemicals, Osaka, Japan)
acetic acid (Wako Pure Chemicals, Osaka, Japan)
ammonia solution (Wako Pure Chemicals, Osaka, Japan)
Partially oxidize a pyridylamino (PA-) sugar chain (0.01 – 1 nmol) with a mixture of 20 μL of 50 mM sodium metaperiodate solution and 20 μL of 0.1 M sodium acetate, pH 4.0 at 0°C for 15 min in the dark.
Add 40 μL of 0.26 M sodium borohydride solution.
Incubate at 25°C for 1 h.
Add 10 μL of acetic acid and mix well.
Neutralize with aqueous ammonia solution.
Analyze the cleaved PA-sugar chain by HPLC as described in the other protocol (Fluorescent labeling of glycans and HPLC analysis).
1) Abdel Akher, M., Hamilton, J., Montgomery, R., Smith, F. (1952) A new procedure for the determination of the fine structure of polysaccharides. J. Am. Chem. Soc. 74 (19) 4970-4971. [PMID : not found]
2) Smith, F., Van Cleve, J. W. (1955) Reduction of the products of periodate oxidation of carbohydrates. I. Hydrogenation with raney nickel of te dialdehydes from the methyl glycopyranosides. J. Am. Chem. Soc. 77 (11) 3091-4971. [PMID : not found]
3) Omichi, K., Hase, S. (1995) Determination of the linkage positions of reducing-end residues of oligosaccharides by partial periodate oxidation of pyridylaminated derivatives. Anal. Biochem. 227 (2) 404-407. [PMID : 7573970]