Human dehydroepiandrosterone sulfotransferase: molecular cloning of cDNA and genomic DNA

doi:10.1016/0009-2797(94)90060-4

Chemico-Biological Interactions

Volume 92, Issues 1–3, June 1994, Pages 145-159

https://doi.org/10.1016/0009-2797(94)90060-4 Get rights and content

Abstract

Human tissues contain at least three well-characterized cytoplasmic sulfotransferase (ST) enzymes, dehydroepiandrosterone (DHEA) ST and two of phenol ST (PST). DHEA ST catalyzes the sulfation of DHEA and other steroids. We cloned and expressed two cDNAs for human liver DHEA ST. The cloning strategy involved the design of PCR primers directed against two conserved domains in ST proteins. These primers were used to generate a specific PCR product that was then used successfully to clone cDNAs for DHEA ST from a human liver cDNA library. Two cDNAs were isolated that were approximately 1.1 and 1.8 kb in length. These two clones had identical open reading frames. Both cDNAs produced enzymatically active DHEA ST protein in a mammalian expression system. Northern blot analysis confirmed the presence of 1.1 and 1.8 kb transcripts in human liver. cDNAs for a number of eukaryotic enzymes have now been cloned, and they share significant sequence homology. These ST cDNAs appear to fall into distinct groups on the basis of amino acid sequences of the proteins that they encode, thus demonstrating that the enzymes comprise a gene superfamily. We have also isolated a genomic clone for human DHEA ST that contains approximately 3 kb of 5′-flanking sequence, exon 1 and 1.7 kb of intron 1. Characterization of the structure and regulatory elements of this gene should help to elucidate mechanisms involved in the regulation of DHEA ST in humans.

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Citation Excerpt :
While the three SULT2 isoforms display overlapping substrate specificity toward different hydroxysteroids such as DHEA and pregnenolone, they exhibit tissue-specific distribution (Falany et al., 2006; Geese and Raftogianis, 2001; Otterness and Weinshilboum, 1994; Thomae et al., 2002). For example, SULT2A1 is expressed mainly in the liver, adrenal glands, and intestine (Otterness and Weinshilboum, 1994; Thomae et al., 2002), whereas SULT2B1b is highly expressed in the placenta, prostate, breast, endometrium, ovary, uterus, small intestine, colon, lung, platelet, brain, and skin (Falany et al., 2006; Geese and Raftogianis, 2001). In contrast, no expression of SULT2B1a protein was detected in any of the tissues examined (Falany and Rohn-Glowacki, 2013).
Pregnenolone and dehydroepiandrosterone (DHEA) are hydroxysteroids that serve as biosynthetic precursors for steroid hormones in human body. SULT2B1b has been reported to be critically involved in the sulfation of pregnenolone and DHEA, particularly in the sex steroid-responsive tissues. The current study was designed to investigate the impact of the genetic polymorphisms of SULT2B1 on the sulfation of DHEA and pregnenolone by SULT2B1b allozymes. Ten SULT2B1b allozymes previously prepared were shown to exhibit differential sulfating activities toward DHEA and pregnenolone in comparison to the wild-type enzyme. Kinetic studies revealed further significant changes in their substrate-binding affinity and catalytic activity toward DHEA and pregnenolone. Taken together, these results indicated clearly a profound effect of SULT2B1 genetic polymorphisms on the sulfating activity of SULT2B1b allozymes toward DHEA and pregnenolone, which may have implications in inter-individual variations in the homeostasis of these two important steroid precursors.
New PCOS-like phenotype in older infertile women of likely autoimmune adrenal etiology with high AMH but low androgens
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Yet, as the table also demonstrates, except for still significantly higher oocyte yields, high-AMH in high-T patients demonstrated complete absence of such differences. Most androgen production in females takes place in ovaries and adrenals, with DHEAS almost exclusively considered an adrenal product [12–14]. One, therefore, can assume that low-DHEAS in association with low-T suggests an adrenal cause for low-T.
How anti-Müllerian hormone (AMH) and testosterone (T) interrelate in infertile women is currently largely unknown. We, therefore, in a retrospective cohort study investigated how infertile women with high-AMH (AMH ≥75th quantile; n = 144) and with normal-AMH (25th–75th quantile; n = 313), stratified for low-T (total testosterone ≤19.0 ng/dL), normal-T (19.0–29.0 ng/dL) and high-T (>29.0 ng/dL) phenotypically behaved. Patient age, follicle stimulating hormone (FSH), dehyroepiandrosterone (DHEA), DHEA sulphate (DHEAS), cortisol (C), adrenocorticotrophic hormone (ACTH), IVF outcomes, as well as inflammatory and immune panels were then compared between groups, with AMH and T as variables. We identified a previously unknown infertile PCOS-like phenotype, characterized by high-AMH but, atypically, low-T, with predisposition toward autoimmunity. It presents with incompatible high-AMH and low-T (<19.0 ng/dL), is restricted to lean PCOS-like patients, presenting delayed for tertiary fertility services. Since also characterized by low DHEAS, low-T is likely of adrenal origina, and consequence of autoimmune adrenal insufficiency since also accompanied by low-C and evidence of autoimmunity. DHEA supplementation in such patients equalizes low- to normal-T and normalizes IVF cycle outcomes. Once recognized, this high-AMH/low-T phenotype is surprisingly common in tertiary fertility centers but, currently, goes unrecognized. Its likely adrenal autoimmune etiology offers interesting new directions for investigations of adrenals control over ovarian function via adrenal androgen production.
Associations between peripheral androgens and cortisol in infertile women
2016, Journal of Steroid Biochemistry and Molecular Biology
Citation Excerpt :
Dehydroepiandrosterone (DHEA) has a short half-life, and is quickly sulfonized via sulfotransferase into its 3β-sulfate, considered its storage form (DHEAS). The SULT2A1 gene primarily codes this conversion in the zona reticularis of the adrenal cortex [13–15], and to lesser degrees in liver and small intestines. Orally ingested DHEA is sulfonized primarily in liver and intestines.
Testosterone has in recent years been proven essential for normal growth and maturation of small growing follicles. Concomitantly, low functional ovarian reserve (LFOR), characterized by a small growing follicle pool, has been associated with low testosterone levels, which can be of ovarian and/or adrenal origin. In this study we, therefore, investigated whether peripheral sex steroid precursors and testosterone levels potentially reflect on adrenal function. In a retrospective cohort study of 355 consecutive infertile women, who presented to an academically affiliated fertility center in New York City, we investigated in a series of statistical models whether low peripheral sex steroid precursors and testosterone are associated with peripheral cortisol (C) levels, reflecting adrenal function. To determine potential correlations, we investigated the dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), androstenedione (AD), total testosterone (TT), free testosterone (FT); sex hormone binding globulin (SHBG), anti-Müllerian hormone (AMH), thyroid stimulating hormone (TSH) and C in a series of multivariate and logistic regression analyses, utilizing C either as a continuous variable or with cut off <5.0 μg/dL, and TT only as a continuous variable. Practically all models demonstrated significant predictability of peripheral sex hormone precursors for C levels, with DHEA demonstrating the strongest and most consistent predictability as an individual parameter and as part of the DHEAS/DHEA ratio. We conclude that in infertile women peripheral sex hormone precursors, especially DHEA, reflect C levels and, therefore, adrenal function. In infertile women, at all ages low levels of sex hormone precursors, therefore, should be considered indications for further adrenal assessments.
Structural plasticity in the human cytosolic sulfotransferase dimer and its role in substrate selectivity and catalysis
2015, Drug Metabolism and Pharmacokinetics
Citation Excerpt :
These sequences provided the platform necessary to begin screening human cDNA libraries for potential human SULTs (hSULTs) [25]. Human cDNA library screening led to the cloning and identification of the first human SULT, an isoform which exhibits activity toward DHEA (hSULT2A1) [27]. Following this discovery, human SULTs with various properties and activities were identified, eventually leading to the current catalog of 14 human SULTs (Table 1).
The cytosolic sulfotransferases (SULTs) are dimeric enzymes that help maintain homeostasis through the modulation of hormone and drug activity by catalyzing their transformation into hydrophilic sulfate esters and increasing their excretion. Each of the thirteen active human SULT isoforms displays a unique substrate specificity pattern that underlies its individual role in our bodies. These specificities have proven to be complex, in some cases masking the biological role of specific isoforms. The first part of this review offers a short summary of historical underpinnings of human SULTs, primarily centered on the characterization of each isoform's kinetic and structural properties. Recent structural investigations have revealed each SULT has an active site “lid” that undergoes restructuring once the cofactor/sulfonate donor, 3′-phosphoadenosine-5′-phosphosulfate (PAPS), binds to the enzyme. This structural rearrangement can alter substrate-binding profiles, therefore complicating enzyme/substrate interactions and making substrate/cosubstrate concentrations and binding order important considerations in enzyme functionality. Molecular dynamic simulations have recently been employed to describe this restructuring in an attempt to offer insight to its effects on substrate selectivity. In addition to reviewing new data on SULT molecular dynamics, we will discuss the contribution of PAPS concentrations and SULT dimerization in the regulation of SULT activity within the human body.
Desulfo-glucosinolate sulfotransferases isolated from several Arabidopsis thaliana ecotypes differ in their sequence and enzyme kinetics
2013, Plant Physiology and Biochemistry
Citation Excerpt :
The varying aa in the ecotypes analyzed in this study are distributed over the whole sequence length (Fig. 2). The structure of SOT proteins in general has been analyzed and four different regions have been defined for either substrate (ds-Gl) or co-substrate (PAPS) binding [19]. The pattern KSGTTW in the N-terminus might be responsible for interaction with 5′-phosphate [20].
The goal was to investigate whether the diverse glucosinolate (Gl) profiles described for different Arabidopsis thaliana (L.) Heynh. ecotypes are at least partially shaped by the kinetic properties of sulfotransferases (SOTs) (EC 2.8.2.-) catalyzing the final step in Gl core structure biosynthesis. This study focuses on only one of the three SOTs that contribute to Gl biosynthesis. Homologues of AtSOT18 proteins were characterized, which was inspired by earlier findings on SOTs from ecotypes Col-0 and C24 differing in two amino acids (aa) and specific enzyme activities. Could there be a correlation of AtSOT18 enzyme activities and differences in Gl profiles between the ecotypes? SOT18 sequences from eight Arabidopsis ecotypes with highly diverse Gl patterns differed in two aa at various positions in the protein sequence. The SOT18 sequence from Col-0 showed the highest similarity to the largest number of other sequences in the alignment. The small differences in the primary sequence lead to important structural changes in secondary and tertiary structure that might be the key of different kinetic activities towards a broad range of substrates. All recombinant AtSOT18 proteins showed low substrate specificity with an indolic Gl, while the specificity for aliphatic substrates varied. There is no correlation in the kinetic behavior with the major ds-Gl contents or with the ratio of C₃/C₄ ds-Gl in the respective ecotype. Therefore, is it unlikely that ds-Gl AtSOT18 proteins play a major role in shaping the Gl profile in Arabidopsis.
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This article is part of a Special Issue entitled: Neuroactive Steroids: Focus on Human Brain.

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Human dehydroepiandrosterone sulfotransferase: molecular cloning of cDNA and genomic DNA

Abstract

Biochim. Biophys. Acta

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Pharmacol. Ther.

Biochem. Biophys. Res. Commun.

Biochim. Biophys. Acta

Adv. Appl. Math.

Purification and characterization of human liver dehydroepiandrosterone sulphotransferase

Biochem. J.

Sulfation of estrone and 17-β estradiol in human liver: catalysis by thermostable phenol sulfotransferase and by dehydroepiandrosterone sulfotransferase

Drug Metab. Dispos.

Human liver steroid sulphotransferase sulphates bile acids

Biochem. J.

Cholesterol sulfation in human liver: catalysis by dehydroepiandrosterone sulfotransferase

Drug Metab. Dispos.

Phenol sulfotransferase in humans: properties, regulation, and function

Hydroxysteroid sulfotransferase and a specific UDP-glucuronosyltransferase are involved in the metabolism of digitoxin in man

Naunyn-Schmiedeberg's Arch. Pharmacol.

Biosynthesis of estrogens in pregnancy: precursor role of plasma dehydroisoandrosterone

Obstet. Gynecol.

A prospective study of dehydroepiandrosterone sulfate, mortality, and cardiovascular disease

N. Engl. J. Med.

Dehydroepiandrosterone sulfate, incidence of myocardial infarction, and extent of atherosclerosis in men

Circulation

Dehydroepiandrosterone (DHEA) levels in patients with prostatic cancer, heart disease and under surgery stress

Exp. Clin. Endocrinol.