Purification and properties of arylsulphatase A from chicken brain.
1. Chicken brain arylsulphatase A was purified 2000-fold, with overall recovery 14%, by using ammonium sulphate fractionation, ethanol precipitation, Sephadex G-200 gel filtration and DEAE-Sephadex column chromatography. 2. The purified preparation was free from beta-glucuronidase, beta-galactosidase, acid phosphatase, inorganic pyrophosphatase and adenosine 3'-phosphate 5'-sulphatophosphate sulphohydrolase activities. 3. Polyacrylamide-gel electrophoresis indicated that the purified preparation was not homogeneous. 4. Chicken brain arylsulphatase was markedly inhibited by carbonyl reagents in the presence of traces of Cu(2+) in the system. Other metal ions such as Fe(2+) and Zn(2+), were inactive. 5. Ascorbic acid alone had no effect on enzyme activity but enhances the inhibition by Cu(2+). 6. Chicken brain arylsulphatase A resembled arylsulphatase A of other animal species in its kinetic properties such as K(m) value, anomalous time-activity relationship and the inhibitory effect of phosphate, sulphite and sulphate ions. However, its electrophoretic mobility, behaviour under zinc acetate fractionation and stimulation by Ag(+) were similar to arylsulphatase B of other animal species. Thus, this enzyme did not correspond to either arylsulphatase A or arylsulphatase B but properties of both. 7. The purified enzyme preparation can degrade cerebroside 3-sulphate.
D000818:Animals; D001205:Ascorbic Acid; D001921:Brain / Q000201:enzymology*; D002645:Chickens; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003300:Copper; D004586:Electrophoresis; D000431:Ethanol; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007501:Iron; D007700:Kinetics; D051381:Rats; D012834:Silver; D013429:Sulfatases / Q000302:isolation & purification*; D013460:Sulfur Isotopes; D015032:Zinc
D013460:Sulfur Isotopes; D000431:Ethanol; D012834:Silver; D003300:Copper; D007501:Iron; D013429:Sulfatases; D015032:Zinc; D001205:Ascorbic Acid