Human fetal adrenal hydroxysteroid sulphotransferase: cDNA cloning, stable expression in V79 cells and functional characterisation of the expressed enzyme.
Dehydroepiandrosterone sulphate (DHEAS) is a major adrenal secretory product, particularly in the fetus where it serves as a substrate for oestrogen biosynthesis by the placenta. The enzyme in the adrenal responsible for synthesising DHEAS, hydroxysteroid sulphotransferase (HST), is therefore essential for human development. We have isolated a full-length cDNA clone, encoding human fetal adrenal HST, and constructed a stable cell line expressing it by transfection into V79 Chinese hamster lung fibroblast cells. This cDNA was essentially identical to that isolated from adult human liver, where the role of HST is less well understood. This recombinant cell line allowed determination of the substrate specificity and kinetic properties of this enzyme towards various steroid hormones, and by comparison of these activities with human liver cytosol we have shown that HST is the major sulphotransferase responsible for the sulphation of DHEA, androsterone and pregnenolone in man and that, functionally, the hepatic and adrenal enzymes are very similar. The expressed HST was also active with testosterone, cortisol (although at low levels) and the xenobiotic 17 alpha-ethinyloestradiol, but not with oestrone or 1-naphthol. We have therefore created a valuable resource for the study of this important enzyme.
Forbes
K J
KJ
Department of Biochemical Medicine, University of Dundee, Ninewells Hospital and Medical School, Scotland, UK.
Hagen
M
M
Glatt
H
H
Hume
R
R
Coughtrie
M W
MW
D000311:Adrenal Glands / Q000196:embryology* / Q000201:enzymology*; D000595:Amino Acid Sequence; D000738:Androsterone / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D018076:DNA, Complementary / Q000235:genetics*; D003687:Dehydroepiandrosterone / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008168:Lung; D008969:Molecular Sequence Data; D011284:Pregnenolone / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D013431:Sulfates / Q000378:metabolism; D015238:Sulfotransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D018076:DNA, Complementary; D011994:Recombinant Proteins; D013431:Sulfates; D003687:Dehydroepiandrosterone; D011284:Pregnenolone; D000738:Androsterone; D015238:Sulfotransferases; C028152:alcohol sulfotransferase