Purification of 3α-hydroxysteroid and 3β-hydroxysteroid dehydrogenases from human liver cytosol

Takikawa, Hajime; Fujiyoshi, Makoto; Nishikawa, Kou; Yamanaka, Masami

doi:10.1002/hep.1840160214

Cited by 12 publications

(7 citation statements)

References 23 publications

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“…This result is in contrast to data of Takikawa et al 22 who describe a 32-kDa bile acid 3␤-HSD from human liver cytosol that does not fit the molecular mass of the ADH monomer of 40 kDa. However, no further information except for molecu- (Table 2).…”

Section: Discussioncontrasting

confidence: 99%

Human liver class I alcohol dehydrogenaseγγ isozyme: the sole cytosolic 3β-hydroxysteroid dehydrogenase of iso bile acids

Marschall¹,

Oppermann²,

Svensson³

et al. 2000

Hepatology

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3beta-Hydroxy (iso) bile acids are formed during enterohepatic circulation from 3alpha-hydroxy bile acids and constitute normal compounds in plasma but are virtually absent in bile. Isoursodeoxycholic acid (isoUDCA) is a major metabolite of UDCA. In a recent study it was found that after administration of isoUDCA, UDCA became the major acid in bile. Thus, epimerization of the 3beta-hydroxy to a 3alpha-hydroxy group, catalyzed by 3beta-hydroxysteroid dehydrogenases (HSD) and 3-oxo-reductases must occur. The present study aims to characterize the human liver bile acid 3beta-HSD. Human liver cytosol and recombinant alcohol dehydrogenase (ADH) betabeta and gammagamma isozymes were subjected to native polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing. Activity staining with oxidized nicotinamide adenine dinucleotide (NAD(+)) or oxidized nicotinamide adenine dinucleotide phosphate (NADP(+)) as cofactors and various iso bile acids as substrates was used to screen for 3beta-HSD activity. Reaction products were identified and quantified by gas chromotography/mass spectrometry (GC/MS). Computer-assisted substrate docking of isoUDCA to the active site of a 3-dimensional model of human class I gammagamma ADH was performed. ADH gammagamma isozyme was identified as the iso bile acid 3beta-HSD present in human liver cytosol, with NAD(+) as a cofactor. Values for k(cat)/K(m) were in the rank order isodeoxycholic acid (isoDCA), isochenodeoxycholic acid (isoCDCA), isoUDCA, and isolithocholic acid (isoLCA) (0.10, 0.09, 0.08, and 0. 05 min(-1) x micromol/L(-1), respectively). IsoUDCA fits as substrate to the 3-dimensional model of the active-site of ADH gammagamma. ADH gammagamma isozyme was defined as the only bile acid 3beta-HSD in human liver cytosol. Hydroxysteroid dehydrogenases are candidates for the binding and transport of 3alpha-hydroxy bile acids. We assume that ADH gammagamma isozyme is involved in cytosolic bile acid binding and transport processes as well.

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Section: Discussioncontrasting

confidence: 99%

Human liver class I alcohol dehydrogenaseγγ isozyme: the sole cytosolic 3β-hydroxysteroid dehydrogenase of iso bile acids

Marschall¹,

Oppermann²,

Svensson³

et al. 2000

Hepatology

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“…There is a significant similarity between the C-3 epimerization by the HSDs and that by the liver cytosol enzyme previously reported [3]; i.e., the coenzyme requirement (NAD and NADPH) and epimerization process (dehydrogenation and reduction). Although HSDs originating from Pseudomonas testasteroni were used in the present study, the main localization of these enzymes in animals and humans is the liver cytosol fraction [18]. These data suggest that some HSDs may catalyze the C-3 epimerization of vitamin D compounds in animals and humans.…”

Section: Resultsmentioning

confidence: 96%

Analysis of C-3 epimerization in (24R)-24,25-dihydroxyvitamin D3 catalyzed by hydroxysteroid dehydrogenase

Higashi

Sakajiri

Shimada

2004

Journal of Pharmaceutical and Biomedical Analysis

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“…3β-HSD was purified from human liver by Takikawa et al [57,58]. The adult mouse liver expresses 3β-HSD types II, III, and IV, with type III predominating.…”

Section: 3β-hsd Isoforms In the Livermentioning

confidence: 99%

Regulation of 3β-Hydroxysteroid Dehydrogenase/Δ5-Δ4 Isomerase: A Review

Rasmussen

Ekstrand

Zamaratskaia

2013

IJMS

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This review focuses on the expression and regulation of 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (3β-HSD), with emphasis on the porcine version. 3β-HSD is often associated with steroidogenesis, but its function in the metabolism of both steroids and xenobiotics is more obscure. Based on currently available literature covering humans, rodents and pigs, this review provides an overview of the present knowledge concerning the regulatory mechanisms for 3β-HSD at all omic levels. The HSD isoenzymes are essential in steroid hormone metabolism, both in the synthesis and degradation of steroids. They display tissue-specific expression and factors influencing their activity, which therefore indicates their tissue-specific responses. 3β-HSD is involved in the synthesis of a number of natural steroid hormones, including progesterone and testosterone, and the hepatic degradation of the pheromone androstenone. In general, a number of signaling and regulatory pathways have been demonstrated to influence 3β-HSD transcription and activity, e.g., JAK-STAT, LH/hCG, ERα, AR, SF-1 and PPARα. The expression and enzymic activity of 3β-HSD are also influenced by external factors, such as dietary composition. Much of the research conducted on porcine 3β-HSD is motivated by its importance for the occurrence of the boar taint phenomenon that results from high concentrations of steroids such as androstenone. This topic is also examined in this review.

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Purification of 3α-hydroxysteroid and 3β-hydroxysteroid dehydrogenases from human liver cytosol

Cited by 12 publications

References 23 publications

Human liver class I alcohol dehydrogenaseγγ isozyme: the sole cytosolic 3β-hydroxysteroid dehydrogenase of iso bile acids

Human liver class I alcohol dehydrogenaseγγ isozyme: the sole cytosolic 3β-hydroxysteroid dehydrogenase of iso bile acids

Analysis of C-3 epimerization in (24R)-24,25-dihydroxyvitamin D3 catalyzed by hydroxysteroid dehydrogenase

Regulation of 3β-Hydroxysteroid Dehydrogenase/Δ5-Δ4 Isomerase: A Review

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