Fig. 8. Endoglucosamine-6-sulfatase activity of expressed Sulfs.

The conditioned medium of CHO cells transfected with the empty vector alone (A and D), HSulf-1 (B and E), or HSulf-2 (C and F) was prepared as described under “Experimental Procedures.” Porcine intestinal heparin (A–C) or shark cartilage chondroitin 6-sulfate (D–F) were incubated with the conditioned medium and then subsequently digested with either a mixture of bacterial heparinases or chondroitinase ABC. The resulting disaccharide fractions were analyzed by as described. The arrows correspond to the elution positions of authentic unsaturated disaccharide markers. The dotted lines indicate the concentrations of KH₂PO₄ used for elution.