A rapid four column purification of 2-deoxy- D-glucoside-2-sulphamate sulphohydrolase from human liver

Abstract

2-Deoxy- D-glucoside-2-sulphamate sulphohydrolase was extracted from human liver and purified 40 000-fold by a simple four column procedure. The purification was followed using a specific substrate isolated from an acid hydrolysate of heparin, O-(α-2- sulphamino-2- deoxy- D-glucopyranosyl )-(1→3)- L-[6, 3H] idonic acid. Only one form of the enzyme was seen on either ion exchange chromatography or isoelectric focussing, with a p I of 6.8. The apparent M r of the haloenzyme as determined by gel filtration was 190 000 ± 20 000. Two other larger M r protein peaks observed on gel filtration appear to be an inactive dimer of the 190 000 dalton peak and a larger aggregate near the exclusion limit of the column. On polyacrylamide disc gel electrophoresis in sodium dodecyl sulphate, with or without prior reduction, each protein peak from the gel filtration column electrophoreced as a single major band with an apparent M r corresponding to 55 000 ± 6000.