Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase with minoxidil as the substrate

Patrick E. Kudlacek, Dahn L Clemens, Christine M. Halgard, Robert J. Anderson

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Biotransformation of xenobiotics and hormones through sulfate conjugation is an important metabolic pathway in humans. The activation of minoxidil, an antihypertensive agent and hair growth stimulator, by sulfation (sulfonation) is carried out by more than one sulfotransferase. Initially only the thermostable form of phenol sulfotransferase was thought to catalyze minoxidil sulfation. We document in this report the new finding that human liver dehydroepiandrosterone sulfotransferase (DHEA ST), an hydroxysteroid sulfotransferase distinct from phenol sulfotransferases, also catalyzes the reaction. To characterize more precisely the activity of DHEA ST toward minoxidil, we used COS-1 cells to express DHEA ST from a human liver cDNA clone. The apparent K(m) values for minoxidil and [35S]3'-phosphoadenosine-5'-phosphosulfate were 3.9 mM and 0.13 μM, respectively. The 50% inactivation temperature of the COS-expressed enzyme was 42°, and the IC50 value for 2,6-dichloro-4-nitrophenol was 1.4 x 10-4M. Both the thermal stability behavior and response to DCNP were similar when the cDNA encoded DHEA ST was assayed with DHEA or minoxidil as a substrate. NaCl led to a greater activation of the cDNA-expressed DHEA ST when assayed with DHEA (2.5-fold) than when the same preparation was assayed with minoxidil (1.4-fold). These data indicate that DHEA ST catalyzes the sulfate conjugation of minoxidil. DHEA ST activity present in the human gut and liver would be expected to add to the overall sulfate conjugation of orally administered minoxidil. Thus, DHEA ST activity must be considered when determining the human tissue sulfotransferase contribution to minoxidil sulfation.

Original languageEnglish (US)
Pages (from-to)215-221
Number of pages7
JournalBiochemical Pharmacology
Volume53
Issue number2
DOIs
StatePublished - Jan 24 1997

Fingerprint

Minoxidil
Liver
Substrates
Sulfonation
Arylsulfotransferase
Sulfotransferases
Sulfates
Dehydroepiandrosterone
Complementary DNA
Phosphoadenosine Phosphosulfate
Chemical activation
dehydroepiandrosterone sulfotransferase
COS Cells
Xenobiotics
Biotransformation
Metabolic Networks and Pathways
Hair
Antihypertensive Agents
Inhibitory Concentration 50
Clone Cells

Keywords

  • antihype rtensive
  • cytosolic hydroxysteroid sulfotransferase
  • dehydroepiandrosterone
  • minoxidil
  • sulfonation
  • sulfotransferase

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

Cite this

Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase with minoxidil as the substrate. / Kudlacek, Patrick E.; Clemens, Dahn L; Halgard, Christine M.; Anderson, Robert J.

In: Biochemical Pharmacology, Vol. 53, No. 2, 24.01.1997, p. 215-221.

Research output: Contribution to journalArticle

Kudlacek, Patrick E. ; Clemens, Dahn L ; Halgard, Christine M. ; Anderson, Robert J. / Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase with minoxidil as the substrate. In: Biochemical Pharmacology. 1997 ; Vol. 53, No. 2. pp. 215-221.
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AB - Biotransformation of xenobiotics and hormones through sulfate conjugation is an important metabolic pathway in humans. The activation of minoxidil, an antihypertensive agent and hair growth stimulator, by sulfation (sulfonation) is carried out by more than one sulfotransferase. Initially only the thermostable form of phenol sulfotransferase was thought to catalyze minoxidil sulfation. We document in this report the new finding that human liver dehydroepiandrosterone sulfotransferase (DHEA ST), an hydroxysteroid sulfotransferase distinct from phenol sulfotransferases, also catalyzes the reaction. To characterize more precisely the activity of DHEA ST toward minoxidil, we used COS-1 cells to express DHEA ST from a human liver cDNA clone. The apparent K(m) values for minoxidil and [35S]3'-phosphoadenosine-5'-phosphosulfate were 3.9 mM and 0.13 μM, respectively. The 50% inactivation temperature of the COS-expressed enzyme was 42°, and the IC50 value for 2,6-dichloro-4-nitrophenol was 1.4 x 10-4M. Both the thermal stability behavior and response to DCNP were similar when the cDNA encoded DHEA ST was assayed with DHEA or minoxidil as a substrate. NaCl led to a greater activation of the cDNA-expressed DHEA ST when assayed with DHEA (2.5-fold) than when the same preparation was assayed with minoxidil (1.4-fold). These data indicate that DHEA ST catalyzes the sulfate conjugation of minoxidil. DHEA ST activity present in the human gut and liver would be expected to add to the overall sulfate conjugation of orally administered minoxidil. Thus, DHEA ST activity must be considered when determining the human tissue sulfotransferase contribution to minoxidil sulfation.

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