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. 2016 Sep 19;291(44):23294–23304. doi: 10.1074/jbc.M116.757328

FIGURE 2.

FIGURE 2.

HPLC analysis of 2-aminobenzamide-labeled disaccharide composition by enzymatic digestion of N2 (wild-type) and RB813 (C41C4.1 mutant) nematodes. HPLC analysis of chondroitinase ABC (CSase) digest of N2 (a), chondroitinase ABC and chondro-4-O-sulfatase (C4Sase) digest of N2 (b), chondroitinase ABC and chondro-6-O-sulfatase (C6Sase) digest of N2 (c), and chondroitinase ABC digest of RB813 (d). a–d, the enzyme digest of the C. elegans GAG fraction was derivatized with 2-aminobenzamide and analyzed by HPLC using an amine-bound silica column (37). The peak eluting at ∼5 min and marked by an asterisk in the inset represents an acetate ion derived from the incubation buffer and an unknown substance derived from the chondroitinase ABC preparation, respectively. The insets show the magnified chromatogram between 30 and 35 min. The elution positions of authentic unsaturated disaccharides labeled with 2-aminobenzamide are indicated by numbered arrows: 1, ΔHexUAα1–3GalNAc; 2, ΔHexUAα1–3GalNAc(4-O-sulfate).