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 s...

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Bibliographic Details
Published inBiochimica et biophysica acta Vol. 657; no. 2; pp. 344 - 355
Main Authors Lim, C T, Horwitz, A L
Format Journal Article
LanguageEnglish
Published Netherlands 13.02.1981
Subjects
Chondroitinases and Chondroitin Lyases - metabolism
Chondroitinsulfatases - isolation & purification
Chondroitinsulfatases - metabolism
Female
Fibroblasts - enzymology
Humans
Kinetics
Placenta - enzymology
Polysaccharides - pharmacology
Pregnancy
Skin - enzymology
Substrate Specificity
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Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:0006-3002
DOI:10.1016/0005-2744(81)90320-X