Endocrine steroid sulfotransferases: Steroid alcohol sulfotransferase from human breast carcinoma cell line MCF-7

https://doi.org/10.1016/0022-4731(86)90331-6Get rights and content

Abstract

Steroid alcohol sulfotransferase (SAS) has been isolated from the cytosol of a human breast carcinoma cell line, MCF-7. This enzyme from Sephadex G-200 chromatography displayed a mol. wt of 118 KDa. The conditions for optimal enzymic activity of SAS were determined to be 20 min incubations at 45°C in 0.2 M Tris buffer (pH 7.5) containing 0.06 M Mg2+. Chromatofocusing chromatography also yielded a single peak of SAS with a pI of 5.8. Results from the incubations of a series of androstane analogues revealed that SAS required a 3β-hydroxyl on a steroid with the trans bridge between the A and B rings. Neither the 3β-allylic hydroxyl group nor the A-ring phenolic 3-hydroxyl accepted the sulfate group from 3'-phosphoadenosine-5'-phosphosulfate. d-ring β-hydroxyl groups were tolerated by the enzyme, however, α-hydroxyl groups on the d-ring appeared to interfere with the reaction. Sulfurylation of steroids by SAS was related inversely to the sum of the displacements of the 3-hydroxyl plus that of the 17-hydroxyl groups relative to the plane of symmetry of the dehydroepiandrosterone nucleus. This enzyme was also capable of sulfurylating short chain aliphatic alcohols, although at greatly reduced rates. 3β-Chloro-5-androstene-17-one and 2-nitroestradiol.17β proved to be the best inhibitors of SAS.

References (37)

  • R.A. Ryan et al.

    Studies on 3β-hydroxy-steroid sulfotransferase from rat liver

    Biochim. biophys. Acta

    (1976)
  • G.W. Oertel et al.

    Effects of steroids on glucose-6-phosphate dehydrogenase

    J. steroid Biochem.

    (1972)
  • R. Gugler et al.

    Reinigung und charakterisierung einer 3'-phosphoadenylylsulfat: steroid sulfotransferase aus der leber des menschen

    Biochim. biophys. Acta

    (1970)
  • J.B. Adams et al.

    Enzymic synthesis of steroid sulfates. XII. Isolation of dehydroepiandrosterone sulfotransferase from human adrenals by affinity chromatography

    Biochim. biophys. Acta

    (1979)
  • E.S. Lyon et al.

    The identity of alcohol sulfotransferases with hydroxysteroid sulfotransferases

    Archs Biochem. Biophys.

    (1980)
  • M. Bouthillier et al.

    The purification of 3β-hydroxysteroid sulfotransferase of the hamster epididymis

    J. steroid Biochem.

    (1985)
  • L. Tseng et al.

    Cyclic change in estradiol metabolic enzymes in human endometrium during the menstrual cycle

  • B.A. Pack et al.

    Nuclear and cytosolic estrogen receptor in gilt endometrium throughout the estrous cycle

    Endocrinology

    (1978)
  • Cited by (0)

    View full text