Heparin with heparanase

Scheme 5.—Typical Cleavage of Heparin (Arbitrary Sequence) with Heparinase (HEPase) and Heparanase (HSase). [Fragments longer than a tetrasaccharide may be obtained by separate use of each enzyme. The combined use of the two enzymes produces only disaccharides.]...

Heparanase and type IV collagenase activities were assessed by a method described previously [17]. Briefly, H-labeled heparin sulfate (HS) was incubated at 37°C with partially purified heparanase in the presence or absence of chitin derivatives. [ H]HS degradation products in the supernatant were analyzed by size-exclusion chromatography. Heparanase activity was determined by measuring the radioactivity of each fraction. H-labeled type IV collagen was incubated at 37°C for 48 h in the presence or absence of chitin derivatives. Type IV collagenolytic activity was calculated from the radioactivity in the supernatant. 

Fig. 8 in a SCM-chitin III concentration-dependent manner. CM-chitin showed no inhibitory effect at a concentration of 500 xg/mL. SCM-chitin III inhibited HS degradation by partially purified heparanase from B16-BL6 cells as did heparin in a concentration-dependent manner (Fig. 9), whereas CM-chitin had little influence at any of the indicated concentrations, except that of 1000 p.g/mL. On the other hand, degradation of H-labeled type IV collagen was inhibited over a 24-h period by more than 75% after treatment with 400 xg/mL of SCM-chitin III and approximately 20% inhibition after treatment with heparin. Little inhibition was shown by CM-chitin. A summary of the biological functions of SCM-chitin III and heparin are listed in Table 5. 

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