Purification and some characteristics of an oestrogen sulphotransferase from guinea pig adrenal gland and its non-identity with adrenal pregnenolone sulphotransferase.
An oestrogen sulphotransferase, active towards both oestrone and oestradiol, and of high specific activity, is present in cytosol prepared from adrenal glands of both sexes of English Shorthair and Hartley guinea pigs. The ovarian and testicular cytosolic activities of this enzyme are markedly low in comparison with the adrenal activity. The adrenal enzyme is distinct from an accompanying pregnenolone sulphotransferase as judged by f.p.l.c. gel filtration, chromatofocusing, and differences in activation brought about by the addition of thiol groups. The oestrogen sulphotransferase behaved as a 67 kDa protein on a Sephadex G100 column and as a 48 kDa protein on f.p.l.c. gel-filtration columns. Two forms of the enzyme with apparent pI values of 6.1 and 5.5 were eluted during f.p.l.c. chromatofocusing. Sequential salt fractionation, f.p.l.c. gel filtration and elution from an agarose-hexane-adenosine-3',5'-diphosphate affinity gel has resulted in a preparation which, when resubmitted to f.p.l.c. gel filtration, yields a considerably purified oestrogen sulphotransferase. When submitted to SDS/polyacrylamide-gel electrophoresis under reducing conditions, a main protein band of 34-36 kDa is observed. It is suggested that the enzyme may exist as a dimer in the cytosol.
Hobkirk
R
R
Department of Biochemistry, University of Western Ontario, London, Canada.
Glasier
M A
MA
Brown
L Y
LY
D000311:Adrenal Glands / Q000201:enzymology*; D000818:Animals; D004967:Estrogens / Q000378:metabolism*; D005260:Female; D006168:Guinea Pigs; D008297:Male; D011284:Pregnenolone / Q000378:metabolism*; D013045:Species Specificity; D013379:Substrate Specificity; D015238:Sulfotransferases; D013466:Sulfurtransferases / Q000302:isolation & purification / Q000378:metabolism*
D004967:Estrogens; D011284:Pregnenolone; D013466:Sulfurtransferases; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; C020122:estrone sulfotransferase