Prokaryotic Expression and Enzyme Activity Measurement of the Human Arylsulfatase (HSulf-1)

The Human arylsulfatase(HSulf-1),which can alter the sulfation state of Heparan Sulfate Proteoglycans(HSPGs) in the neutral condition,is a secreted protein.The shift of sulfation state of Heparan Sulfate Proteoglycans has the affection on the combination of the signaling molecule and its receptor thereby has regulated the signaling pathway.Given the potential values of HSulf-1's application,in order to gain sufficient protein to develop its function and application research,the functional fragment of HSulf-1 is constructed into expression vector,pGEX-6p-1 to form a recombinant plasmid.Expression,isolation,and purification of the fragment of functional domain of Hsulf-1 in the prokaryotic expression system are performed.Then,the enzyme activity of prokaryotic expression product is measured by the method of fluorospectrophotometry using the 4-Mus as substrate.The data suggest that the purified protein can be obtained through the prokaryotic expression system but have no enzyme activity.