Purification of a soluble adenosine 5′-phosphosulphate sulphohydrolase from bovine liver

Abstract

Studies of the subcellular distribution of adenosine 5′-phosphosulphate (APS) sulphohydrolase have indicated a bimodal distribution of the enzyme between the lysosomal and supernatant fractions of the bovine liver cell. The supernatant enzyme has been purified 1200-fold by ammonium sulphate fractionation, DEAE-cellulose chromatography and fractionation on Sephadex G-100. Samples of the preparation gave a single homegeneous enzymically active component on polyacrylamide disc gel electrophoresis. The molecular weight was 68 000–69 000 by sodium dodecyl sulphate -polyacrylamide gel electrophoresis and by gel filtration on Sephadex G-100. The enzyme was specific for APS and was free of 3′-phosphoadenosine 5′-phosphosulphate sulphohydrolase and 3′-nucleotidase activities.