Differential xenoestrogen-sulfating activities of the human cytosolic sulfotransferases: molecular cloning, expression, and purification of human SULT2B1a and SULT2B1b sulfotransferases

Ethanol Sulfation by the Human Cytosolic Sulfotransferases: A Systematic Analysis

“…It should be pointed out that, in contrast to a previous study on ethanol sulfation that had been performed using recombinant human SULTs expressed in Salmonella typhimurium, 16) purified recombinant human SULTs, with purities greater than 95% as judged by SDS-PAGE, [23][24][25][26] were used in the present study.…”

“…A metabolic labeling study using HepG2 human hepatoma cells labeled with [ Preparation of Purified Human SULTs Recombinant human P-form (SULT1A1 and SULT1A2) and Mform (SULT1A3) phenol SULTs, thyroid hormone SULT (SULT1B1), two SULT1Cs (SULT1C2 and SULT1C4), estrogen SULT (SULT1E1), dehydroepiandrosterone (DHEA) SULT (SULT2A1), and a neuronal SULT (SULT4A1), expressed using pGEX-2TK prokaryotic expression system, and two SULT2B1s (designated a and b) expressed using pET23c expression system, were prepared and purified as previously described. [23][24][25][26] Preparation of HepG2 Cell Lysate Confluent HepG2 cells grown in a 25 cm 2 culture flask were lysed in 300 µL of a lysis buffer containing 1% (w/w) NP-40, 0.02 M potassium phosphate buffer, pH 7.5, 0.15 M NaCl, 5 mM EDTA, 50 mM sodium fluoride, and protease inhibitor cocktail. HepG2 cell lysate prepared was cleared by centrifugation at 25000×g for 30 min at 4°C, and the supernatant collected was used in the SULT assay.…”

Phase Ii Drug Metabolizing Enzymes

“…Other substrates include norepinephrine, catechols, monocyclic phenols and aromatic molecules 50 . The substrate specifity of SULT1B1 is restricted to thyroid hormones 60 and small phenolic compounds such as 1-naphtol and 4-nitrophenol 61 . SULT1C1 conjugates some iodothyronines 62 but a good substrate for this enzyme has not been identified.…”

On the Sulfation and Methylation of Catecholestrogens in Human Mammary Epithelial Cells and Breast Cancer Cells

“…[27][28][29][30] Enzymatic Assay The sulfating activity of the recombinant human SULTs was assayed using PAP[ Control with DMSO alone was also prepared. The reaction was started by the addition of the enzyme (1.25 mg), allowed to proceed for 10 min at 37°C, and stopped by placing the assay mixture-containing thin-wall tube on a heating block, pre-heated to 100°C, for 2 min.…”