Human cytosolic steroid sulfotransferases: Versatile and rapid activity assays

Abstract

Conjugation of steroids and sterol compounds with a sulfonate group is a major pathway in the regulation of their activity, synthesis and excretion. Three human cytosolic sulfotransferases are highly involved in the sulfonation of sterol compounds. SULT1E1 has a low nM affinity for estrogen sulfonation and also conjugates non-aromatic steroids with a significantly lower affinity. SULT2A1 is responsible for the high levels of fetal and adult dehydroepiandrosterone (DHEA) sulfate synthesis in the adrenal gland as well as many 3α and 3ß-hydroxysteroids and bile acids. SULT2B1b is responsible for the majority of cholesterol sulfation in tissues as well as conjugating 3ß-hydroxysteroids. Although there are multiple methods for assaying cytosolic SULT activity, two relatively simple, rapid and versatile assays for steroid sulfonation are described. The first method utilizes radiolabeled substrates and organic solvent extraction to isolate the radiolabeled product from the aqueous phase. The second assay utilizes ³⁵S-3'-phosphoadenosine 5'-phosphosulfate (PAPS) to generate ³⁵S-conjugated products that are resolved by thin layer chromatography. Both assays useful in situations requiring measurement of SULT activity in a timely manner.

Keywords: 3′,5’-diphosphoadenosine (PAP); 3′-phosphoadenosine 5′-phosphosulfate (PAPS); dehydroepiandrosterone (DHEA); phase 2 conjugation; sulfonation; sulfotransferase (SULT); ß-Estradiol (ß-E2).