Immunochemical characterisation of a dehydroepiandrosterone sulfotransferase in rats and humans.
A member of the rat liver hydroxysteroid sulfotransferase (ST) enzyme family metabolising dehydroepiandrosterone (DHEA) was purified from female rats and used to raise rabbit polyclonal antibodies. Characterisation of this antibody preparation demonstrated that it was specific for DHEA ST, and recognised a single 30-kDa protein on immunoblot analysis of rat liver cytosol which was expressed preferentially in female rat liver, and immunohistochemical localisation of the protein in female rat liver determined that DHEA ST was distributed homogeneously in the cytoplasm of hepatocytes. Examination of the extrahepatic expression of this protein showed it to be located predominantly in the liver, although a small amount of enzyme activity was found in the kidney which was not apparently subject to the same sex difference as the hepatic activity. Immunological analysis suggested that this activity was not due to the action of DHEA ST, but to another, unidentified ST isozyme. The antibody cross-reacted strongly with adult human liver DHEA ST, recognising a protein of 35 kDa on immunoblotting. Using this antibody preparation, the distribution of DHEA ST in mid-trimester human fetal tissues was examined, and it was shown that the enzyme is expressed in the adrenal and liver, but not to any significant extent in the kidney or lung. This antibody therefore provides a powerful tool for investigating the function of DHEA ST.
Sharp
S
S
Department of Biochemical Medicine, University of Dundee, Ninewells Hospital and Medical School, Scotland.
Barker
E V
EV
Coughtrie
M W
MW
Lowenstein
P R
PR
Hume
R
R
D000311:Adrenal Glands / Q000196:embryology / Q000201:enzymology; D000818:Animals; D000906:Antibodies; D003687:Dehydroepiandrosterone / Q000378:metabolism*; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G; D007150:Immunohistochemistry; D008099:Liver / Q000196:embryology / Q000201:enzymology; D008297:Male; D009928:Organ Specificity; D051381:Rats; D017208:Rats, Wistar; D015238:Sulfotransferases / Q000032:analysis* / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D007074:Immunoglobulin G; D003687:Dehydroepiandrosterone; D015238:Sulfotransferases; C019998:dehydroepiandrosterone sulfotransferase