Journal article
Bacterial Expression, Purification, and Characterization of Rat Hydroxysteroid Sulfotransferase STa
Protein expression and purification, Vol.21(1), pp.235-242
02/2001
DOI: 10.1006/prep.2000.1364
PMID: 11162411
Abstract
Hydroxysteroid (alcohol) sulfotransferase catalyzes numerous reactions that are important to our understanding of the metabolism of both endogenous steroids and exogenous alcohols. Here we report a method for prokaryotic expression and rapid purification of the recombinant hydroxysteroid sulfotransferase STa, a major isoform of hydroxysteroid sulfotransferase in the rat. The cDNA encoding STa was cloned into a pET-3c vector and expressed in Escherichia coli BL21 cells. After disruption of the cells by sonication, the enzyme was purified in one step by affinity chromatography on adenosine 3′,5′-diphosphate–agarose. The purified recombinant STa had a relative molecular mass on SDS–PAGE that was identical with the native hepatic STa in rat liver. The expressed enzyme displayed similar substrate inhibition characteristics with dehydroepiandrosterone as have been noted previously with the native enzyme purified from rat liver. Furthermore, the catalytic efficiency in sulfation of 7-hydroxymethyl-12-methylbenz[a]anthracene, as well as the stereoselectivity of sulfation of the enantiomers of 1-phenyl-1-heptanol and 1-naphthyl-1-ethanol, catalyzed by the recombinant STa were consistent with characteristics of the STa isolated from rat liver.
Details
- Title: Subtitle
- Bacterial Expression, Purification, and Characterization of Rat Hydroxysteroid Sulfotransferase STa
- Creators
- Jonathan J. Sheng - Division of Medicinal and Natural Products Chemistry, College of Pharmacy, Iowa City, IA 52242, USA.Michael W. Duffel - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Protein expression and purification, Vol.21(1), pp.235-242
- Publisher
- Elsevier Inc
- DOI
- 10.1006/prep.2000.1364
- PMID
- 11162411
- ISSN
- 1046-5928
- eISSN
- 1096-0279
- Language
- English
- Date published
- 02/2001
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984303162602771
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