Cited by 224Estrogen and related hormone effectsPharmacogenetics and Drug MetabolismChemical Reaction Mechanisms

Sulfuryl transfer: the catalytic mechanism of human estrogen sulfotransferase

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Zhang, Huiping, Varmalova, Olga, Vargas, Froyland M., Falany, Charles N., Leyh, Thomas S.·1998·Journal of Biological Chemistry

Abstract

Estrogen sulfotransferase (EST) catalyzes the transfer of the sulfuryl group from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to 17beta-estradiol (E2). The sulfation of E2 prevents it from binding to, and thereby activating, the estrogen receptor. The regulation of EST appears to be causally linked to tumorigenesis in the breast and endometrium. In this study, recombinant human EST is characterized, and the catalytic mechanism of the transfer reaction is investigated in ligand binding and initial rate experiments. The native enzyme is a dimer of 35-kDa subunits. The apparent equilibrium constant for transfer to E2 is (4.5 +/- 0.2) x 10(3) at pH 6.3 and T = 25 +/- 2 degrees C. Initial rate studies provide the kinetic constants for the reaction and suggest a sequential mechanism. E2 is a partial substrate inhibitor (Ki = 80 +/- 5 nM). The binding of two E2 per EST subunit suggests that the partial inhibition occurs through binding at an allosteric site. In addition to providing the dissociation constants for the ligand-enzyme complexes, binding studies demonstrate that each substrate binds independently to the enzyme and that both the E.PAP.E2S and E.PAP.E2 dead-end complexes form. These results strongly suggest a Random Bi Bi mechanism with two dead-end complexes.

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Crossref · 10.1074/jbc.273.18.10888

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  1. Falany, J. L., and Falany, C. N. (1998) Oncol. Res., in press

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Authors
Zhang, H., Varmalova, O., Vargas, F. M., Falany, C. N., & Leyh, T. S.
Affiliations
  • Albert Einstein College of Medicine (US)
  • University of Alabama at Birmingham (US)
Corresponding author
Leyh, Thomas S.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1998-05
Volume
273
Issue
18
Pages
10888-10892
ISSN
0021-9258 (ISSN-L)
OpenAlex
W1968379731
Cited by
223
References
26
Citations by year
2026: 3 · 2025: 5 · 2024: 5 · 2023: 4 · 2022: 5 · 2021: 5 · 2020: 4 · 2019: 4 · 2018: 6 · 2017: 3 · 2016: 7 · 2015: 10 · 2014: 13 · 2013: 12 · 2012: 15
FWCI
3.82
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (GM54469, GM38953)
  • National Institute of General Medical Sciences (GM 54469, R01GM054469, GM-38953)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
  • Physical Sciences › Chemistry › Organic Chemistry › Chemical Reaction Mechanisms
Keywords
estrogen sulfotransferase, PAPS, catalytic mechanism, sequential mechanism, dead-end complex, substrate inhibition, allosteric binding, enzyme kinetics, estrogen receptor, breast cancer, endometrial cancer, enzyme dimer
MeSH terms
Catalysis, Cell Line, Chromatography, Gel, Humans, Hydrogen-Ion Concentration, Kinetics, Molecular Weight, Protein Binding, Sulfites, Sulfotransferases

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