Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase (DHEA ST) with minoxidil as the substrate

P. Kudlacek, D. Clemens, C. Halgard, R. Anderson

Research output: Contribution to journalArticle

Abstract

Minoxidil (MNX) is a potent antihypertensive and hair growth promoter. Vasodilatory and hair follicle effects are due to MNX-sulfate, the active metabolite produced by both thermostable and thermolabile phenol sulfotransferases. Because our initial work indicated that DHEA ST might also catalyze MNX sulfation, we tested and characterized human liver cDNA expressed DHEA ST activity with MNX. Apparent Km values for MNX and 3′ -phosphoadenosine-5′ -phosphosulfate were 3. 9 mM and 0. 13 μ M, respectively. Thermal stability of the cDNA encoded DHEA ST activity with MNX and DHEA as substrates was nearly identical as indicated by a 50% inactivation temperature of 42° C for both. The IC50 values for activity assayed with MNX and DHEA were 1. 4 × 10-4 M and 6. 0 × 105 M, respectively. Activity assayed with MNX and DHEA in the presence of NaCI showed activation of 1. 4-fold at 25 mM NaCI and 2. 5-fold at 200 mM NaCI, respectively. HPLC of the reaction product showed that 99% of the net radioactivity coeluted with authentic MNX-sulfate. Conclusion: DHEA ST, an hydroxysteroid ST, catalyzes the sulfation of MNX and must be considered when determining the contribution of human tissue sulfotransferases to MNX sulfation. (Supported, in part, by the VA Medical Research Service and Creighton University).

Original languageEnglish (US)
Pages (from-to)A456
JournalFASEB Journal
Volume10
Issue number3
StatePublished - Dec 1 1996

Fingerprint

Minoxidil
sulfotransferases
prasterone
Liver
liver
Substrates
Dehydroepiandrosterone
sulfates
inactivation temperature
Arylsulfotransferase
Complementary DNA
Phosphoadenosine Phosphosulfate
Hydroxysteroids
hair follicles
antihypertensive agents
biomedical research
Hair Follicle
dehydroepiandrosterone sulfotransferase
Sulfotransferases
thermal stability

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase (DHEA ST) with minoxidil as the substrate. / Kudlacek, P.; Clemens, D.; Halgard, C.; Anderson, R.

In: FASEB Journal, Vol. 10, No. 3, 01.12.1996, p. A456.

Research output: Contribution to journalArticle

@article{dac567aa15284a6fa00505260337f381,
title = "Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase (DHEA ST) with minoxidil as the substrate",
abstract = "Minoxidil (MNX) is a potent antihypertensive and hair growth promoter. Vasodilatory and hair follicle effects are due to MNX-sulfate, the active metabolite produced by both thermostable and thermolabile phenol sulfotransferases. Because our initial work indicated that DHEA ST might also catalyze MNX sulfation, we tested and characterized human liver cDNA expressed DHEA ST activity with MNX. Apparent Km values for MNX and 3′ -phosphoadenosine-5′ -phosphosulfate were 3. 9 mM and 0. 13 μ M, respectively. Thermal stability of the cDNA encoded DHEA ST activity with MNX and DHEA as substrates was nearly identical as indicated by a 50{\%} inactivation temperature of 42° C for both. The IC50 values for activity assayed with MNX and DHEA were 1. 4 × 10-4 M and 6. 0 × 105 M, respectively. Activity assayed with MNX and DHEA in the presence of NaCI showed activation of 1. 4-fold at 25 mM NaCI and 2. 5-fold at 200 mM NaCI, respectively. HPLC of the reaction product showed that 99{\%} of the net radioactivity coeluted with authentic MNX-sulfate. Conclusion: DHEA ST, an hydroxysteroid ST, catalyzes the sulfation of MNX and must be considered when determining the contribution of human tissue sulfotransferases to MNX sulfation. (Supported, in part, by the VA Medical Research Service and Creighton University).",
author = "P. Kudlacek and D. Clemens and C. Halgard and R. Anderson",
year = "1996",
month = "12",
day = "1",
language = "English (US)",
volume = "10",
pages = "A456",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "3",

}

TY - JOUR

T1 - Characterization of recombinant human liver dehydroepiandrosterone sulfotransferase (DHEA ST) with minoxidil as the substrate

AU - Kudlacek, P.

AU - Clemens, D.

AU - Halgard, C.

AU - Anderson, R.

PY - 1996/12/1

Y1 - 1996/12/1

N2 - Minoxidil (MNX) is a potent antihypertensive and hair growth promoter. Vasodilatory and hair follicle effects are due to MNX-sulfate, the active metabolite produced by both thermostable and thermolabile phenol sulfotransferases. Because our initial work indicated that DHEA ST might also catalyze MNX sulfation, we tested and characterized human liver cDNA expressed DHEA ST activity with MNX. Apparent Km values for MNX and 3′ -phosphoadenosine-5′ -phosphosulfate were 3. 9 mM and 0. 13 μ M, respectively. Thermal stability of the cDNA encoded DHEA ST activity with MNX and DHEA as substrates was nearly identical as indicated by a 50% inactivation temperature of 42° C for both. The IC50 values for activity assayed with MNX and DHEA were 1. 4 × 10-4 M and 6. 0 × 105 M, respectively. Activity assayed with MNX and DHEA in the presence of NaCI showed activation of 1. 4-fold at 25 mM NaCI and 2. 5-fold at 200 mM NaCI, respectively. HPLC of the reaction product showed that 99% of the net radioactivity coeluted with authentic MNX-sulfate. Conclusion: DHEA ST, an hydroxysteroid ST, catalyzes the sulfation of MNX and must be considered when determining the contribution of human tissue sulfotransferases to MNX sulfation. (Supported, in part, by the VA Medical Research Service and Creighton University).

AB - Minoxidil (MNX) is a potent antihypertensive and hair growth promoter. Vasodilatory and hair follicle effects are due to MNX-sulfate, the active metabolite produced by both thermostable and thermolabile phenol sulfotransferases. Because our initial work indicated that DHEA ST might also catalyze MNX sulfation, we tested and characterized human liver cDNA expressed DHEA ST activity with MNX. Apparent Km values for MNX and 3′ -phosphoadenosine-5′ -phosphosulfate were 3. 9 mM and 0. 13 μ M, respectively. Thermal stability of the cDNA encoded DHEA ST activity with MNX and DHEA as substrates was nearly identical as indicated by a 50% inactivation temperature of 42° C for both. The IC50 values for activity assayed with MNX and DHEA were 1. 4 × 10-4 M and 6. 0 × 105 M, respectively. Activity assayed with MNX and DHEA in the presence of NaCI showed activation of 1. 4-fold at 25 mM NaCI and 2. 5-fold at 200 mM NaCI, respectively. HPLC of the reaction product showed that 99% of the net radioactivity coeluted with authentic MNX-sulfate. Conclusion: DHEA ST, an hydroxysteroid ST, catalyzes the sulfation of MNX and must be considered when determining the contribution of human tissue sulfotransferases to MNX sulfation. (Supported, in part, by the VA Medical Research Service and Creighton University).

UR - http://www.scopus.com/inward/record.url?scp=33749142067&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749142067&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:33749142067

VL - 10

SP - A456

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 3

ER -