Substrate gating confers steroid specificity to estrogen sulfotransferase

Evgeniy V. Petrotchenko, Mary E. Doerflein, Yoshimitsu Kakuta, Lars C. Pedersen, Masahiko Negishi

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Estrogen sulfotransferase (EST) exhibits a high substrate specificity and catalytic efficiency toward estrogens such as estradiol (E2) but insignificant ability to sulfate hydroxysteroids such as dehydroepiandrosterone (DHEA). To provide the structural basis for this estrogen specificity, we mutated amino acid residues that constitute the substrate-binding site of EST. Among these mutants, only Tyr-81 decreased E2 and increased DHEA sulfotransferase activities. Substitution for Tyr-81 by smaller hydrophobic residues increased K(m(E2)) for E2 activity, whereas the k(cat(E2)) remained relatively constant. The Y81L mutant exhibited the same DHEA activity as wild-type hydroxysteroid sulfotransferase, for which K(m(DHEA)) remained relatively constant, and k(cat(DHEA)) was markedly increased. The side chain of Tyr-81 is directed at the A-ring of the E2 molecule in the substrate-binding pocket of EST, constituting a steric gate with Phe-142 sandwiching E2 from the opposite side. The present mutagenesis study indicates that the 3β-hydroxyl group of the DHEA molecule is excluded from the catalytic site of EST through steric hindrance of Tyr-81 with the C- 19 methyl group of DHEA. Thus, this stricture-like gating caused by steric hindrance appears to be a structural principle for conferring estrogen specificity to EST.

Original languageEnglish
Pages (from-to)30019-30022
Number of pages4
JournalJournal of Biological Chemistry
Volume274
Issue number42
DOIs
Publication statusPublished - Oct 15 1999
Externally publishedYes

Fingerprint

Dehydroepiandrosterone
Steroids
Substrates
Estrogens
Cats
Hydroxysteroids
Mutagenesis
Molecules
Substrate Specificity
Hydroxyl Radical
Sulfates
estrone sulfotransferase
Estradiol
Catalytic Domain
Pathologic Constriction
Substitution reactions
Binding Sites
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Petrotchenko, E. V., Doerflein, M. E., Kakuta, Y., Pedersen, L. C., & Negishi, M. (1999). Substrate gating confers steroid specificity to estrogen sulfotransferase. Journal of Biological Chemistry, 274(42), 30019-30022. https://doi.org/10.1074/jbc.274.42.30019

Substrate gating confers steroid specificity to estrogen sulfotransferase. / Petrotchenko, Evgeniy V.; Doerflein, Mary E.; Kakuta, Yoshimitsu; Pedersen, Lars C.; Negishi, Masahiko.

In: Journal of Biological Chemistry, Vol. 274, No. 42, 15.10.1999, p. 30019-30022.

Research output: Contribution to journalArticle

Petrotchenko, EV, Doerflein, ME, Kakuta, Y, Pedersen, LC & Negishi, M 1999, 'Substrate gating confers steroid specificity to estrogen sulfotransferase', Journal of Biological Chemistry, vol. 274, no. 42, pp. 30019-30022. https://doi.org/10.1074/jbc.274.42.30019
Petrotchenko, Evgeniy V. ; Doerflein, Mary E. ; Kakuta, Yoshimitsu ; Pedersen, Lars C. ; Negishi, Masahiko. / Substrate gating confers steroid specificity to estrogen sulfotransferase. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 42. pp. 30019-30022.
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