Bacterial expression and characterization of a cDNA for human liver estrogen sulfotransferase

J Steroid Biochem Mol Biol. 1995 Jun;52(6):529-39. doi: 10.1016/0960-0760(95)00015-r.


A distinct human estrogen sulfotransferase (hEST-1) cDNA has been isolated from a human liver lambda Zap cDNA library using a PCR procedure. The enzymatically active protein has been expressed in two bacterial expression systems and the kinetic and immunologic properties of the enzyme have been characterized. The full-length cDNA for hEST-1 is 994 base pairs in length and encodes a 294 amino acid protein with a calculated molecular mass of 35,123 Da. Purified hEST-1 migrated with an apparent molecular mass of 35,000 Da during SDS-polyacrylamide gel electrophoresis. Immunoblot analysis of hEST-1 expressed in E. coli with a rabbit anti-hEST-1 antibody yields a band of approximately 35,000 Da. The anti-hEST-1 antibody also detects a single band in human liver and jejunum cytosol which migrates with the same molecular mass as expressed hEST-1. There was also no cross-reactivity of hEST-1 with rabbit anti-hP-PST or rabbit anti-hDHEA-ST antibodies upon immunoblot analysis. hEST-1 was expressed in bacteria and purified to homogeneity. Expressed hEST-1 activity has a significantly greater affinity for estrogen sulfation than that found for the other human STs which conjugate estrogens. hEST-1 maximally sulfates beta-estradiol and estrone at concentrations of 20 nM. hEST-1 also sulfates dehydroepiandrosterone, pregnenolone, ethinylestradiol, and 1-naphthol, at significantly higher concentrations; however, cortisol, testosterone and dopamine are not sulfated. The results presented in this paper describe the expression and characterization of a human EST distinct from other human STs which sulfate estrogens. The high affinity of hEST-1 for estrogens indicates that this ST may be important in both the metabolism of estrogens and in the regulation of their activities.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • DNA, Complementary / genetics*
  • Escherichia coli / genetics
  • Estrogens / metabolism
  • Gene Expression
  • Humans
  • Kinetics
  • Liver / enzymology*
  • Molecular Sequence Data
  • Molecular Weight
  • Polymerase Chain Reaction
  • Rabbits
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Substrate Specificity
  • Sulfotransferases / genetics*
  • Sulfotransferases / immunology
  • Sulfotransferases / metabolism


  • DNA, Complementary
  • Estrogens
  • Sulfotransferases
  • estrone sulfotransferase

Associated data

  • GENBANK/S77383