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. 1989 Jun 15;260(3):641–646. doi: 10.1042/bj2600641

Purification and characterization of human liver dehydroepiandrosterone sulphotransferase.

C N Falany 1, M E Vazquez 1, J M Kalb 1
PMCID: PMC1138726  PMID: 2764897

Abstract

A form of sulphotransferase capable of sulphating dehydroepiandrosterone and other steroids was purified from cytosol prepared from human liver. Dehydroepiandrosterone sulphotransferase was purified 621-fold when compared with the activity in cytosol using DEAE-Sepharose CL-6B and adenosine 3',5'-bisphosphate-agarose affinity chromatography. During affinity chromatography, dehydroepiandrosterone sulphation activity could be resolved from p-nitrophenol sulphation activity catalysed by phenol sulphotransferase by using a gradient of adenosine 3'-phosphate 5'-phosphosulphate. The purified enzyme was most active towards dehydroepiandrosterone but was capable of conjugating a number of other steroids, including pregnenolone, androsterone and beta-oestradiol. No activity towards p-nitrophenol or dopamine, substrates for the phenol sulphotransferase, was observed with the pure enzyme. A single band with a subunit molecular mass of 35 kDa was observed by Coomassie Blue staining following SDS/polyacrylamide-gel electrophoresis of the purified enzyme. A molecular mass of 68-70 kDa was calculated for the active form of the enzyme by chromatography on Sephacryl S-200, suggesting that the active form of the enzyme is a dimer.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams J. B., McDonald D. Enzymic synthesis of steroid sulphates. XII. Isolation of dehydroepiandrosterone sulphotransferase from human adrenals by affinity chromatography. Biochim Biophys Acta. 1979 Mar 16;567(1):144–153. doi: 10.1016/0005-2744(79)90181-5. [DOI] [PubMed] [Google Scholar]
  2. Adams J. B., McDonald D. Enzymic synthesis of steroid sulphates. XIII. Isolation and properties of dehydroepiandrosterone sulphotransferase from human foetal adrenals. Biochim Biophys Acta. 1980 Sep 9;615(1):275–278. doi: 10.1016/0005-2744(80)90031-5. [DOI] [PubMed] [Google Scholar]
  3. Adams J. B., McDonald D. Enzymic synthesis of steroid sulphates. XIV. Properties of human adrenal steroid alcohol sulphotransferase. Biochim Biophys Acta. 1981 Jun 23;664(3):460–468. doi: 10.1016/0005-2760(81)90124-7. [DOI] [PubMed] [Google Scholar]
  4. Barańczyk-Kuźma A. Phenol sulfotransferase in human lung. Biochem Med Metab Biol. 1986 Feb;35(1):18–30. doi: 10.1016/0885-4505(86)90054-x. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  6. Campbell N. R., Van Loon J. A., Weinshilboum R. M. Human liver phenol sulfotransferase: assay conditions, biochemical properties and partial purification of isozymes of the thermostable form. Biochem Pharmacol. 1987 May 1;36(9):1435–1446. doi: 10.1016/0006-2952(87)90108-0. [DOI] [PubMed] [Google Scholar]
  7. Chen L. J., Segel I. H. Purification and characterization of bile salt sulfotransferase from human liver. Arch Biochem Biophys. 1985 Sep;241(2):371–379. doi: 10.1016/0003-9861(85)90559-4. [DOI] [PubMed] [Google Scholar]
  8. Falany C. N., Green M. D., Swain E., Tephly T. R. Substrate specificity and characterization of rat liver p-nitrophenol, 3 alpha-hydroxysteroid and 17 beta-hydroxysteroid UDP-glucuronosyltransferases. Biochem J. 1986 Aug 15;238(1):65–73. doi: 10.1042/bj2380065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Falany C. N., Tephly T. R. Separation, purification and characterization of three isoenzymes of UDP-glucuronyltransferase from rat liver microsomes. Arch Biochem Biophys. 1983 Nov;227(1):248–258. doi: 10.1016/0003-9861(83)90368-5. [DOI] [PubMed] [Google Scholar]
  10. Foldes A., Meek J. L. Rat brain phenolsulfotransferase: partial purification and some properties. Biochim Biophys Acta. 1973 Dec 19;327(2):365–374. doi: 10.1016/0005-2744(73)90419-1. [DOI] [PubMed] [Google Scholar]
  11. Gordon G. B., Shantz L. M., Talalay P. Modulation of growth, differentiation and carcinogenesis by dehydroepiandrosterone. Adv Enzyme Regul. 1987;26:355–382. doi: 10.1016/0065-2571(87)90023-9. [DOI] [PubMed] [Google Scholar]
  12. Gugler R., Rao G. S., Breuer H. Reinigung und Charakterisierung einer 3'-Phosphoadenylylsulfat: Steroid-Sulfotransferase aus der Leber des Menschen. Biochim Biophys Acta. 1970 Oct 14;220(1):69–84. doi: 10.1016/0005-2744(70)90230-5. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Marcus C. J., Sekura R. D., Jakoby W. B. A hydroxysteroid sulfotransferase from rat liver. Anal Biochem. 1980 Sep 15;107(2):296–304. doi: 10.1016/0003-2697(80)90386-3. [DOI] [PubMed] [Google Scholar]
  15. Reiter C., Mwaluko G., Dunnette J., Van Loon J., Weinshilboum R. Thermolabile and thermostable human platelet phenol sulfotransferase. Substrate specificity and physical separation. Naunyn Schmiedebergs Arch Pharmacol. 1983 Sep;324(2):140–147. doi: 10.1007/BF00497020. [DOI] [PubMed] [Google Scholar]
  16. Sekura R. D., Duffel M. W., Jakoby W. B. Aryl sulfotransferases. Methods Enzymol. 1981;77:197–206. doi: 10.1016/s0076-6879(81)77026-5. [DOI] [PubMed] [Google Scholar]
  17. Singer S. S., Giera D., Johnson J., Sylvester S. Enzymatic sulfation of steroids: I. The enzymatic basis for the sex difference in cortisol sulfation by rat liver preparations. Endocrinology. 1976 Apr;98(4):963–974. doi: 10.1210/endo-98-4-963. [DOI] [PubMed] [Google Scholar]
  18. Sugiyama Y., Stolz A., Sugimoto M., Kuhlenkamp J., Yamada T., Kaplowitz N. Identification and partial purification of a unique phenolic steroid sulphotransferase in rat liver cytosol. Biochem J. 1984 Dec 15;224(3):947–953. doi: 10.1042/bj2240947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Whittemore R. M., Roth J. A. A modified Ecteola cellulose assay for M and P phenol sulfotransferase. Biochem Pharmacol. 1985 May 15;34(10):1647–1652. doi: 10.1016/0006-2952(85)90629-x. [DOI] [PubMed] [Google Scholar]

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