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. 2014 Aug 13;289(40):27886–27898. doi: 10.1074/jbc.M114.590752

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Sequencing analysis of the HCLase-resistant tetrasaccharide fraction. A, the purity of the HCLase-resistant tetrasaccharide fraction from CS-D was confirmed through 2-AB labeling followed by anion-exchange HPLC. B, the disaccharide composition of the resistant tetrasaccharide was determined by comparing the elution positions of the 2-AB-labeled unsaturated disaccharides produced by digestion using CSase ABC with those of authentic 2-AB-derivatized unsaturated CS disaccharides indicated by arrows. C, the disaccharide moiety at the reducing end of the resistant tetrasaccharide was identified by digesting the 2-AB-labeled tetrasaccharides with a novel CSase from Vibrio sp. FC509. All samples were analyzed by HPLC on a YMC-Pack Polyamine II column using a NaH₂PO₄ gradient (indicated by the dashed line). 1, Δ^4,5HexUAα1-3GalNAc(6S); 2, Δ^4,5HexUA(2S)α1-3GalNAc(6S); 3, the resistant tetrasaccharide.