TABLE 3.

Enzymatic activity of the hHYAL4 mutants generated by introducing mutation(s) in domain I of h(R305Q)

Nonlabeled CS-A was incubated individually with the purified protein (125 ng each) at 37 °C in 50 mm acetate buffer, pH 4.5, for 30 min. The hydrolytic activity of the construct h(I)(R305Q) toward CS-A is taken as 100%. All constructs having the hydrolytic activity exhibited the mouse-type specificity.

Constructs	Relative activity
h(I)(R305Q)	100
h(S261A,I262V,G263S,V264I,W265R)(R305Q)	23
h(S261A,I262V,G263S)(R305Q)	6
h(I262V,G263S,V264I)(R305Q)	4
h(G263S,V264I,W265R)(R305Q)	16
h(S261A,I262V)(R305Q)	NDa
h(G263S)(R305Q)	ND
h(V264I,W265R)(R305Q)	ND
h(L268F,G269A)(R305Q)	ND
h(R305Q)	ND

^a ND, not detected.