Monte Carlo-minimized energy profile of estradiol in the ligand-binding tunnel of 17?-hydroxysteroid dehydrogenase: Atomic mechanisms of steroid recognition

Zhorov, Boris S.; Lin, Szu-Yuan

doi:10.1002/(sici)1097-0134(20000301)38:4<414::aid-prot7>3.0.co;2-x

Cited by 22 publications

(17 citation statements)

References 41 publications

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“…The active site extends to a proper size to accommodate the substrate when a steroid approaches the site. This is in agreement with the recent results in the study of other steroid-converting enzymes, such as the significant active site volume modification in binding different steroids found in the type 5 human 17␤-hydroxysteroid dehydrogenase (HSD), 2 and the conformational rearrangements found when estradiol advances in the binding site of human estrogenic 17␤-HSD (31).…”

Section: Resultssupporting

confidence: 92%

Identifying Androsterone (ADT) as a Cognate Substrate for Human Dehydroepiandrosterone Sulfotransferase (DHEA-ST) Important for Steroid Homeostasis

Chang

Shi

Rehse

et al. 2004

Journal of Biological Chemistry

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In steroid biosynthesis, human dehydroepiandrosterone sulfotransferase (DHEA-ST) in the adrenals has been reported to catalyze the transfer of the sulfonate group from 3-phosphoadenosine-5-phosphosulfate to dehydroepiandrosterone (DHEA). DHEA and its sulfate play roles as steroid precursors; however, the role of the enzyme in the catabolism of androgens is poorly understood. Androsterone sulfate is clinically recognized as one of the major androgen metabolites found in urine. Here it is demonstrated that this enzyme recognizes androsterone (ADT) as a cognate substrate with similar kinetics but a 2-fold specificity and stronger substrate inhibition than DHEA. The structure of human DHEA-ST in complex with ADT has been solved at 2.7 Å resolution, confirming ADT recognition. Structural analysis has revealed the binding mode of ADT differs from that of DHEA, despite the similarity of the overall structure between the ADT and the DHEA binary complexes. Our results identify that this human enzyme is an ADT sulfotransferase as well as a DHEA sulfotransferase, implying an important role in steroid homeostasis for the adrenals and liver.Sulfonation is catalyzed by a family of sulfotransferases that conjugate a sulfonate group (SO 3 ) from 3Ј-phosphoadenosine-5Ј-phosphosulfate (PAPS) 1 to a hydroxyl group of the recipient molecule. With desulfation by sulfatases, sulfonation has been considered as one of the major enzymatic reactions in the metabolism not only of endogenous compounds and xenobiotics, but also of steroid hormones. In most cases, the transfer of the charged sulfonate moiety to an acceptor steroid decreases the biological activity of the steroid. Indeed, steroid sulfates resulting from this reaction are not capable of binding to or activating steroid receptors. In addition, the sulfonation reaction increases water solubility of steroids and thereby enhances their excretion into the urine and/or bile (1, 2).Human dehydroepiandrosterone sulfotransferase (DHEA-ST; SULT2A1; EC 2.8.2.2) was identified mainly from human liver and adrenals, using Northern blot analysis (3) and RT-PCR analysis (4). A single isoform of DHEA-ST from human liver and adrenal tissues was confirmed by the expression and purification of the enzyme from these organs (5, 6), molecular cloning studies (7) and the comparison study of the physical, kinetic, and immunological properties of liver and adrenal forms of the enzyme (8). Steroid sulfonation has been recognized as an important means for maintaining steroid hormone levels in their metabolism. In humans, dehydroepiandrosterone sulfate (DHEAS) is the most prodigious steroid precursor and one of the major secretory products of both adult and fetal adrenals. In the fetoplacental-maternal unit (the unique interdependence of fetus, placenta, and mother) shown in Scheme 1, DHEAS plays an important role as the major precursor for placental estrogen biosynthesis, thus maintaining pregnancy. A considerable amount of DHEAS is mainly produced from the fetal zone in the adrenal gland (9). Then DHEAS...

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Section: Resultssupporting

confidence: 92%

Identifying Androsterone (ADT) as a Cognate Substrate for Human Dehydroepiandrosterone Sulfotransferase (DHEA-ST) Important for Steroid Homeostasis

Chang

Shi

Rehse

et al. 2004

Journal of Biological Chemistry

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“…These mechanisms of selectivity over C19 steroids, steric hindrance at the A-ring and hydrogen bonding of the hydroxyl groups at either end of the molecule, are also seen in the binding of E 2 to ERa . Modelling has also confirmed that cofactor and substrate may bind in either order (Zhorov & Lin 2000), as was seen in early kinetic activity studies (Betz 1971).…”

Section: Inhibitors Of 17b-hsd1supporting

confidence: 60%

Design and validation of specific inhibitors of 17 -hydroxysteroid dehydrogenases for therapeutic application in breast and prostate cancer, and in endometriosis

Day¹,

Tutill²,

Purohit³

et al. 2008

Endocrine Related Cancer

114

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Abstract17b-Hydroxysteroid dehydrogenases (17b-HSDs) are enzymes that are responsible for reduction or oxidation of hormones, fatty acids and bile acids in vivo, regulating the amount of the active form that is available to bind to its cognate receptor. All require NAD(P)(H) for activity. Fifteen 17b-HSDs have been identified to date, and with one exception, 17b-HSD type 5 (17b-HSD5), an aldo-keto reductase, they are all short-chain dehydrogenases/reductases, although overall homology between the enzymes is low. Although named as 17b-HSDs, reflecting the major redox activity at the 17b-position of the steroid, the activities of these 15 enzymes vary, with several of the 17b-HSDs able to reduce and/or oxidise multiple substrates at various positions. These activities are involved in the progression of a number of diseases, including those related to steroid metabolism. Despite the success of inhibitors of steroidogenic enzymes in the clinic, such as those of aromatase and steroid sulphatase, the development of inhibitors of 17b-HSDs is at a relatively early stage, as at present none have yet reached clinical trials. However, many groups are now working on inhibitors specific for several of these enzymes for the treatment of steroid-dependent diseases, including breast and prostate cancer, and endometriosis, with demonstrable efficacy in in vivo disease models. In this review, the recent advances in the validation of these enzymes as targets for the treatment of these diseases, with emphasis on 17b-HSD1, 3 and 5, the development of specific inhibitors, the models used for their evaluation, and their progress towards the clinic will be discussed.

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“…In addition to the energy components described in Ref. 33, the energy expression included hydration energy calculated by the method of Lazaridis and Karplus (34) and energy of deformation of bond angles in testosterone. The enzyme was represented by a two-shell model based on our crystallographic structure.…”

Section: Methodsmentioning

confidence: 99%

“…The MCM energy profile was computed with the help of the ZMM program described in Ref. 33 and references therein.…”

Section: Methodsmentioning

confidence: 99%

Structure of the Human 3α-Hydroxysteroid Dehydrogenase Type 3 in Complex with Testosterone and NADP at 1.25-Å Resolution

Nahoum¹,

Gangloff²,

Legrand³

et al. 2001

Journal of Biological Chemistry

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The first crystallographic structure of human type 3 3␣-hydroxysteroid dehydrogenase (3␣-HSD3, AKR1C2), an enzyme playing a critical role in steroid hormone metabolism, has been determined in complex with testosterone and NADP at 1.25-Å resolution. The enzyme's 17␤-HSD activity was studied in comparison with its 3␣-HSD activity. The enzyme catalyzes the inactivation of dihydrotestosterone into 5␣-androstane-3␣,17␤-diol (3␣-diol) as well as the transformation of androstenedione into testosterone. Using our homogeneous and highly active enzyme preparation, we have obtained 150-fold higher 3␣-HSD specificity as compared with the former reports in the literature. Although the rat and the human 3␣-HSDs share 81% sequence homology, our structure reveals significantly different geometries of the active sites. Substitution of the Ser 222 by a histidine in the human enzyme may compel the steroid to adopt a different binding to that previously described for the rat (Bennett, M. J., Albert, R. H., Jez, J. M., Ma, H., Penning, T. M., and Lewis, M. (1997) Structure 5, 799 -812). Furthermore, we showed that the affinity for the cofactor is higher in the human 3␣-HSD3 than the rat enzyme due to the presence of additional hydrogen bonds on the adenine moiety and that the cofactor is present under its reduced form in the active site in our preparation.Human 3␣-hydroxysteroid dehydrogenases (3␣-HSDs; 1 EC 1.1.1.213) work in concert with the 5␣/5␤-steroid reductases to convert steroid hormones into the 3␣/5␣ and 3␣/5␤-tetrahydrosteroids. These isozymes catalyze the inactivation of androgens, estrogens, progestins, and glucocorticoids. However, the inactivation of the most potent androgen 5␣-dihydrotestosterone (5␣-DHT) to 5␣-androstane-3␣,17␤-diol (3␣-diol) is its best known function (1). These isoenzymes thus play a major role in the regulation of the intracellular concentration of 3␣-DHT in peripheral tissues, especially in the androgen-sensitive prostate that is susceptible to benign prostatic hyperplasia and prostate cancer. Testosterone, after entering prostatic cells, is transformed to 5␣-DHT by 3-oxo-5␣-steroid-4-dehydrogenase (2, 3). 5␣-DHT is a more potent androgen than testosterone in stimulating prostate cancer growth (4, 5) and preferentially binds to the androgen receptor (dissociation constant (K d ) for the androgen receptor of 10 Ϫ11 M) (6). Elevation of 5␣-DHT content in prostate has been associated with benign prostatic hyperplasia in humans (6, 7) and with human prostate carcinoma (8, 9). The action of 5␣-DHT may be terminated by 3␣-HSD, which catalyzes the inactivation of 5␣-DHT to 3␣-androstanediol (a weak androgen; K d for the androgen receptor of 10 Ϫ6 M) (1). It has also been proposed that, by catalyzing the reverse reaction, 3␣-HSD may function as a molecular switch and, in this manner, may regulate the amount of 5␣-DHT available for androgen receptor binding and activation. Three isoforms of 3␣-HSD have been described in human tissues (10 -12) as playing critical roles in sex hormone metabolism and action...

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Monte Carlo-minimized energy profile of estradiol in the ligand-binding tunnel of 17?-hydroxysteroid dehydrogenase: Atomic mechanisms of steroid recognition

Cited by 22 publications

References 41 publications

Identifying Androsterone (ADT) as a Cognate Substrate for Human Dehydroepiandrosterone Sulfotransferase (DHEA-ST) Important for Steroid Homeostasis

Identifying Androsterone (ADT) as a Cognate Substrate for Human Dehydroepiandrosterone Sulfotransferase (DHEA-ST) Important for Steroid Homeostasis

Design and validation of specific inhibitors of 17 -hydroxysteroid dehydrogenases for therapeutic application in breast and prostate cancer, and in endometriosis

Structure of the Human 3α-Hydroxysteroid Dehydrogenase Type 3 in Complex with Testosterone and NADP at 1.25-Å Resolution

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