Investigation of action pattern of a novel chondroitin sulfate/dermatan sulfate 4-O-endosulfatase

Recently, a novel CS/DS 4-O-endosulfatase was identified from a marine bacterium and its catalytic mechanism was investigated further (Wang, W., et. al (2015) J. Biol. Chem. 290, 7823-7832; Wang, S., et. al (2019) Front. Microbiol. 10, 1309). In the study herein, we provide new insight about the structural characteristics of the substrate which determine the activity of this enzyme. The substrate specificities of the 4-O-endosulfatase were probed by using libraries of structure-defined CS/DS oligosaccharides issued from synthetic and enzymatic sources. We found that this 4-O-endosulfatase effectively remove the 4-0-sulfate of disaccharide sequences GlcUA beta 1-3GalNAc(4S) or GlcUA beta 1-3GalNAc(4S,6S) in all tested hexasaccharides. The sulfated GalNac residue is resistant to the enzyme when adjacent uronic residues are sulfated as shown by the lack of enzymatic desulfation of GlcUA-Investigation of action pattern of a novel chondroitin sulfate/dermatan sulfate 4-O-endosulfatase1-3GaINAc(4S) connected to a disaccharide GlcUA(2S)beta 1-3GalNAc(6S) in an octasaccharide. The 3-O-sulfation of GlcUA was also shown to hinder the action of this enzyme. The 4-O-endosulfatase exhibited an oriented action from the reducing to the non-reducing whatever the saturation or not of the non-reducing end. Finally, the activity of the 4-O-endosulfatase decreases with the increase in substrate size. With the deeper understanding of this novel 4-O-endosulfatase, such chondroitin sulfate (CS)/dermatan sulfate (DS) sulfatase is a useful tool for exploring the structure-function relationship of CS/DS.

Automated summary

This study identified a novel CS/DS 4-O-endosulfatase from a marine bacterium and investigated its catalytic mechanism, revealing substrate specificities, action patterns, and a decrease in enzyme activity with increasing substrate size. The deeper understanding of this enzyme provides a useful tool for exploring the structure-function relationship of CS/DS.