The application of chondro-2-sulfatase for identification of the products generated from chondroitin sulfate isomers by high-performance liquid chromatography.

A specific chondroitin sulfate-lyase, chondro-2-sulfatase, was first used for identification of the unsaturated disaccharide constituents (delta Di-S) generated from variously sulfated chondroitin sulfate and dermatan sulfate isomers by a high-performance liquid chromatographic (HPLC) method. delta Di-S generated from oversulfated chondroitin sulfate and dermatan sulfate isomers following digestion with chondroitinases were further digested by the chondro-2-sulfatase, which led to the release of one sulfate from a specific 2-position of the uronic acid residue, as judged with the new HPLC system using a resin made from a sulfonized styrene-divinylbenzene copolymer. It was also found that the chondro-2-sulfatase digests not only delta Di-S with the structure of D-uronic acid 2 sulfate 1-3-N-acetyl-D-galactosamine but also other sulfated delta Di-S with partially the same constituents, i.e., unsaturated di-sulfated disaccharide B, unsaturated di-sulfated disaccharide D or G, and unsaturated tri-sulfated disaccharide.