Purification and properties of human N-acetylgalactosamine-6-sulfate sulfatase.
1. Human N-acetylgalactosamine-6-sulfate sulfatase (EC 3.1.6.-) from human placenta has been purified more than 3000-fold by gel filtration, ion-exchange and substrate affinity chromatography. The enzyme has a molecular weight of 90 000 by gel filtration chromatography and 85 000 by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Enzyme purified from cultured human skin fibroblasts has similar properties. 2. The tritium-labeled chrondroitin 6-sulfate trisaccharide N-acetylgalactosamine 6-sulfate-(beta, 1-4)-glucuronic acid-(beta, 1-3(-N-acetyl[1-3H]galactosaminitol 6-sulfate as substrate demonstrated a Km of 0.12 mM at pH 4.5. Sulfate was hydrolyzed only from the non-reducing terminal of this disulfated trisaccharide. Hyaluronic acid, dermatan sulfate, chondroitin 4-sulfate, heparin and chondroitin 6-sulfate tetrasaccharide were slightly inhibitory, whereas 6-sulfated pentasaccharides and heptasaccharides were strongly inhibitory. The enzyme dose not hydrolyze sulfate from N-acetylglucosamine 6-sulfate.
D002810:Chondroitinases and Chondroitin Lyases / Q000378:metabolism*; D002811:Chondroitinsulfatases / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D010920:Placenta / Q000201:enzymology*; D011134:Polysaccharides / Q000494:pharmacology; D011247:Pregnancy; D012867:Skin / Q000201:enzymology; D013379:Substrate Specificity
D011134:Polysaccharides; D002811:Chondroitinsulfatases; D002810:Chondroitinases and Chondroitin Lyases