Purification and characterization of human liver dehydroepiandrosterone sulphotransferase.
A form of sulphotransferase capable of sulphating dehydroepiandrosterone and other steroids was purified from cytosol prepared from human liver. Dehydroepiandrosterone sulphotransferase was purified 621-fold when compared with the activity in cytosol using DEAE-Sepharose CL-6B and adenosine 3',5'-bisphosphate-agarose affinity chromatography. During affinity chromatography, dehydroepiandrosterone sulphation activity could be resolved from p-nitrophenol sulphation activity catalysed by phenol sulphotransferase by using a gradient of adenosine 3'-phosphate 5'-phosphosulphate. The purified enzyme was most active towards dehydroepiandrosterone but was capable of conjugating a number of other steroids, including pregnenolone, androsterone and beta-oestradiol. No activity towards p-nitrophenol or dopamine, substrates for the phenol sulphotransferase, was observed with the pure enzyme. A single band with a subunit molecular mass of 35 kDa was observed by Coomassie Blue staining following SDS/polyacrylamide-gel electrophoresis of the purified enzyme. A molecular mass of 68-70 kDa was calculated for the active form of the enzyme by chromatography on Sephacryl S-200, suggesting that the active form of the enzyme is a dimer.
Department of Pharmacology and Cancer Center, University of Rochester, NY 14642.
C N
CN
Falany
M E
ME
Vazquez
J M
JM
Kalb
D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D006801:Humans; D008099:Liver / Q000201:enzymology*; D015238:Sulfotransferases / Q000302:isolation & purification* / Q000378:metabolism
D015238:Sulfotransferases; C019998:dehydroepiandrosterone sulfotransferase