Studies of a novel 3-0 sulfatase have been extended. This enzyme is specific for a unique 3-0,N disulfated glucosamine residue in heparin which is essential for its role as an anticoagulant. The purified enzyme provides a useful probe for revealing the presence of this unique derivative in fragments of heparin and heparin like oligosaccharides. The sulfatase has been demonstrated in plasma and in extracts of leukocytes and has been purified approximately 100-fold from human urine. To further the purification, controlled periodate oxidation of heparin has been carried out to produce fragments terminating in sulfated glucosamine residues. These fragments have been linked to sepharose to provide an affinity adsorbant which should prove useful in purifying this enzyme and others of the Sanfilippo complex which act on terminal glucosamine derivatives.
