Estrogen sulfotransferase of mouse placenta: behaviour and characteristics during partial purification

Abstract

Mouse placental estrogen sulfotransferase (ST) was partially purified by fast protein liquid chromatography (FPLC) gel filtration in combination with FPLC anion exchange. Owing to the highly unstable nature of the enzyme, large increases in specific activity were not obtained. Storage of the ST in the presence of thiol groups at -20 degrees C stabilized the enzyme considerably. Forty-three percent of the cytosolic ST was bound to an Affi-Gel blue column and eluted as a broad peak at approximately 0.8 M NaCl. The use of the latter procedure, in combination with FPLC gel filtration, did not increase the specific activity substantially. Larger increases in specific activity were obtained using agarose-hexane-adenosine-3',5'-diphosphate affinity chromatography. The bound ST activity was eluted under a single peak at 1 mM ADP. Increases in specific activity following use of this column averaged 54-fold but could reach 90-fold. Attempts at further purification of this material resulted in low recovery and decreased specific activity. Velocity versus substrate concentration curves show that estrone and particularly estradiol inhibit the partially purified mouse placental sulfotransferase above 0.1-0.25 microM substrate concentrations.

Purification and some characteristics of an oestrogen sulphotransferase from guinea pig adrenal gland and its non-identity with adrenal pregnenolone sulphotransferase

“…Another publication reports a molecular mass of 31 kDa for an unstable oestrogen sulphotransferase from porcine uterus [23]. It has also been demonstrated that the corresponding enzyme in mouse placenta and uterus behaves as if it were a 29-30 kDa protein [7,8]. The presence of two forms of guinea pig adrenal oestrogen sulphotransferase during chromatofocusing parallels information obtained for the mouse placental enzyme [7] and for the guinea pig chorionic enzyme [10].…”

“…The importance of these enzymes in this latter respect has been particularly noted in the mammalian intrauterine compartment [3][4][5][6]. Severe difficulties have been encountered in purifying and characterizing the poorly abundant oestrogen sulphotransferase(s) of intrauterine origin in small rodents including mouse [7,8] and guinea pig [9,10] during gestation. The reported presence of a hydroxysteroid sulphotransferase, which is particularly active toward pregnenolone [11], in the male Hartley guinea pig adrenal cortex, led us to enquire into the possible occurrence of an oestrogen sulphotransferase in that tissue, especially since the bovine adrenal gland contains an abundance of a stable enzyme of this type [12].…”