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William L Duax - One of the best experts on this subject based on the ideXlab platform.
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Divergent evolution of a Rossmann Fold and identification of its oldest surviving ancestor
International journal of bioinformatics research and applications, 2009Co-Authors: William L Duax, Robert Huether, Vladimir Z. Pletnev, Timothy C. Umland, Charles M. WeeksAbstract:β-ketoacyl (acyl carrier protein) reductase (β-k-ACPR) enzymes are essential to fatty acid synthesis in bacteria. The analyses revealed the most primitive member of the β-k-ACPRs family was a NADP reductase where NADP was recognised by a Thr residue in the β2α3 turn. Aromatic residue stacking at the dimer interface and a previously undetected conserved sequence at the C-terminus, stabilise the oligomeric assembly of these proteins. Our analysis indicates that the primordial members of the β-k-ACPR family probably arose in the α-proteobacteria and are characterised by the presence of multiple open reading frames and an extreme codon and amino acid bias.
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structure function of human type 1 3β hydroxysteroid dehydrogenase an intrasubunit disulfide bond in the Rossmann Fold domain and a cys residue in the active site are critical for substrate and coenzyme utilization
The Journal of Steroid Biochemistry and Molecular Biology, 2007Co-Authors: James L Thomas, Robert Huether, Vance L Mack, Launa A Scaccia, Ryan C Stoner, William L DuaxAbstract:The human type 1 (placenta, breast tumors) and type 2 (gonads, adrenals) isoforms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) are key enzymes in steroidogenic pathways leading to the production of all active steroid hormones. Kinetic analyses of purified 3beta-HSD1 show that the Michaelis-Menten constants (Km) for substrates and cofactor are decreased dramatically (three- to eight-Fold) by the addition of beta-mercaptoethanol (BME), which suggest that a disulfide bond may be critical to ligand utilization. Western immunoblots and SDS-PAGE of purified 3beta-HSD1 in the presence or absence of BME showed a lack of intersubunit disulfide bonds in the dimeric enzyme. The Rossmann-Fold domain of 3beta-HSD1 contains two Cys residues, Cys72 and Cys111, which are capable of forming an intrasubunit disulfide bond based on their proximity in our structural model. Our structural model also predicts that Cys83 may affect the orientation of substrate and cofactor. To test these predictions, the C72S, C72F, C111S, C111A, C83S and C83A mutants of 3beta-HSD1 were produced, expressed, and purified. BME failed to diminish the Km values of substrate and cofactor for C72S, C72F, C111S and C111A but produced a 2.5 decrease in Km values for C83A ligands similar to wild-type 3beta-HSD. Thus, our results support the presence of an intrasubunit disulfide bond between Cys72 and Cys111 that participates in the tertiary structure of the Rossmann-Fold domain. Although C83S had no enzyme activity, the C83A mutant enzyme exhibited two- to five-Fold higher Km values for substrate and cofactor but had similar K(cat) values compared to wild-type 3beta-HSD. These data characterize the roles of Cys residues in 3beta-HSD and validate the predictions of our structural model.
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Structure/function of human type 1 3beta-hydroxysteroid dehydrogenase: An intrasubunit disulfide bond in the Rossmann-Fold domain and a Cys residue in the active site are critical for substrate and coenzyme utilization.
The Journal of steroid biochemistry and molecular biology, 2007Co-Authors: James L Thomas, Robert Huether, Vance L Mack, Launa A Scaccia, Ryan C Stoner, William L DuaxAbstract:The human type 1 (placenta, breast tumors) and type 2 (gonads, adrenals) isoforms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) are key enzymes in steroidogenic pathways leading to the production of all active steroid hormones. Kinetic analyses of purified 3beta-HSD1 show that the Michaelis-Menten constants (Km) for substrates and cofactor are decreased dramatically (three- to eight-Fold) by the addition of beta-mercaptoethanol (BME), which suggest that a disulfide bond may be critical to ligand utilization. Western immunoblots and SDS-PAGE of purified 3beta-HSD1 in the presence or absence of BME showed a lack of intersubunit disulfide bonds in the dimeric enzyme. The Rossmann-Fold domain of 3beta-HSD1 contains two Cys residues, Cys72 and Cys111, which are capable of forming an intrasubunit disulfide bond based on their proximity in our structural model. Our structural model also predicts that Cys83 may affect the orientation of substrate and cofactor. To test these predictions, the C72S, C72F, C111S, C111A, C83S and C83A mutants of 3beta-HSD1 were produced, expressed, and purified. BME failed to diminish the Km values of substrate and cofactor for C72S, C72F, C111S and C111A but produced a 2.5 decrease in Km values for C83A ligands similar to wild-type 3beta-HSD. Thus, our results support the presence of an intrasubunit disulfide bond between Cys72 and Cys111 that participates in the tertiary structure of the Rossmann-Fold domain. Although C83S had no enzyme activity, the C83A mutant enzyme exhibited two- to five-Fold higher Km values for substrate and cofactor but had similar K(cat) values compared to wild-type 3beta-HSD. These data characterize the roles of Cys residues in 3beta-HSD and validate the predictions of our structural model.
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Structure/function of human type 1 3β-hydroxysteroid dehydrogenase: An intrasubunit disulfide bond in the Rossmann-Fold domain and a Cys residue in the active site are critical for substrate and coenzyme utilization
The Journal of Steroid Biochemistry and Molecular Biology, 2007Co-Authors: James L Thomas, Robert Huether, Vance L Mack, Launa A Scaccia, Ryan C Stoner, William L DuaxAbstract:The human type 1 (placenta, breast tumors) and type 2 (gonads, adrenals) isoforms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) are key enzymes in steroidogenic pathways leading to the production of all active steroid hormones. Kinetic analyses of purified 3beta-HSD1 show that the Michaelis-Menten constants (Km) for substrates and cofactor are decreased dramatically (three- to eight-Fold) by the addition of beta-mercaptoethanol (BME), which suggest that a disulfide bond may be critical to ligand utilization. Western immunoblots and SDS-PAGE of purified 3beta-HSD1 in the presence or absence of BME showed a lack of intersubunit disulfide bonds in the dimeric enzyme. The Rossmann-Fold domain of 3beta-HSD1 contains two Cys residues, Cys72 and Cys111, which are capable of forming an intrasubunit disulfide bond based on their proximity in our structural model. Our structural model also predicts that Cys83 may affect the orientation of substrate and cofactor. To test these predictions, the C72S, C72F, C111S, C111A, C83S and C83A mutants of 3beta-HSD1 were produced, expressed, and purified. BME failed to diminish the Km values of substrate and cofactor for C72S, C72F, C111S and C111A but produced a 2.5 decrease in Km values for C83A ligands similar to wild-type 3beta-HSD. Thus, our results support the presence of an intrasubunit disulfide bond between Cys72 and Cys111 that participates in the tertiary structure of the Rossmann-Fold domain. Although C83S had no enzyme activity, the C83A mutant enzyme exhibited two- to five-Fold higher Km values for substrate and cofactor but had similar K(cat) values compared to wild-type 3beta-HSD. These data characterize the roles of Cys residues in 3beta-HSD and validate the predictions of our structural model.
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Structure and mechanism of action and inhibition of steroid dehydrogenase enzymes involved in hypertension.
Endocrine research, 1998Co-Authors: William L Duax, Debashis GhoshAbstract:Members of the NADPH-dependent short chain dehydrogenase/reductase (SDR) family control blood pressure, fertility, and natural and neoplastic growth. Despite the fact that only one amino acid residue is strictly conserved in the 100 known members of the family, all appear to have a dinucleotide-binding Rossmann Fold and homologous catalytic residues including the conserved tyrosine. Variation in the binding pocket creates specificity for steroids, prostaglandins, sugars and alcohols. The critically important tyrosine appears to maintain a fixed position relative to the scafFolding of the Rossmann Fold and the cofactor position, while the substrate-binding pocket alters in such a way that the dehydrogenation/reduction reaction site is brought into bonding distance of the tyrosine hydroxyl group. Licorice induces high blood pressure by inhibiting an SDR in the kidney. The crystal structure of the complex of 3α, 20s-hydroxysteroid dehydrogenase and carbenoxolone reveals the mechanism of enzyme inhibition by ...
Robert E. Royer - One of the best experts on this subject based on the ideXlab platform.
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17-beta-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold.
Chemico-biological interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L Vander JagtAbstract:17-beta-Hydroxysteroid dehydrogenase type 1 (17betaHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-beta-estradiol. 17betaHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17betaHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17betaHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17betaHSD1 is a target for identification of lead compounds for structure-based drug design. 17betaHSD1 was purified from human placenta. 17betaHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 microM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17betaHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17betaHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
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17-β-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold
Chemico-Biological Interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L. Vander JagtAbstract:Abstract 17-β-Hydroxysteroid dehydrogenase type 1 (17βHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-β-estradiol. 17βHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17βHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17βHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17βHSD1 is a target for identification of lead compounds for structure-based drug design. 17βHSD1 was purified from human placenta. 17βHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 μM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17βHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17βHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
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Gossypol: prototype of inhibitors targeted to dinucleotide Folds.
Current medicinal chemistry, 2000Co-Authors: David L. Vander Jagt, Lorraine M. Deck, Robert E. RoyerAbstract:Gossypol, a disesquiterpene from cottonseed, exhibits multiple biological properties, including male antifertility activity and anticancer activity. Gossypol also inhibits the growth of numerous parasitic organisms and shows antiviral activity against a number of enveloped viruses, including the AIDS virus. Derivatives of gossypol, in which the aldehyde functional groups that contribute to toxicity have been modified, retain or even show enhanced biological activity. Ring substituted 2,3-dihydroxy-1-naphthoic acids, which are structural analogs of gossypol, share with gossypol the ability to complex with dehydrogenases at the dinucleotide Fold (Rossmann Fold) with selectivity, suggesting that gossypol may be considered the prototype of a new class of drugs targeted to dehydrogenases. Most of the biological activities of gossypol and related compounds may result from inhibition of dehydrogenases.
David L. Vander Jagt - One of the best experts on this subject based on the ideXlab platform.
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17-β-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold
Chemico-Biological Interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L. Vander JagtAbstract:Abstract 17-β-Hydroxysteroid dehydrogenase type 1 (17βHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-β-estradiol. 17βHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17βHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17βHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17βHSD1 is a target for identification of lead compounds for structure-based drug design. 17βHSD1 was purified from human placenta. 17βHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 μM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17βHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17βHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
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Gossypol: prototype of inhibitors targeted to dinucleotide Folds.
Current medicinal chemistry, 2000Co-Authors: David L. Vander Jagt, Lorraine M. Deck, Robert E. RoyerAbstract:Gossypol, a disesquiterpene from cottonseed, exhibits multiple biological properties, including male antifertility activity and anticancer activity. Gossypol also inhibits the growth of numerous parasitic organisms and shows antiviral activity against a number of enveloped viruses, including the AIDS virus. Derivatives of gossypol, in which the aldehyde functional groups that contribute to toxicity have been modified, retain or even show enhanced biological activity. Ring substituted 2,3-dihydroxy-1-naphthoic acids, which are structural analogs of gossypol, share with gossypol the ability to complex with dehydrogenases at the dinucleotide Fold (Rossmann Fold) with selectivity, suggesting that gossypol may be considered the prototype of a new class of drugs targeted to dehydrogenases. Most of the biological activities of gossypol and related compounds may result from inhibition of dehydrogenases.
Lorraine M. Deck - One of the best experts on this subject based on the ideXlab platform.
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17-beta-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold.
Chemico-biological interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L Vander JagtAbstract:17-beta-Hydroxysteroid dehydrogenase type 1 (17betaHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-beta-estradiol. 17betaHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17betaHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17betaHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17betaHSD1 is a target for identification of lead compounds for structure-based drug design. 17betaHSD1 was purified from human placenta. 17betaHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 microM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17betaHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17betaHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
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17-β-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold
Chemico-Biological Interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L. Vander JagtAbstract:Abstract 17-β-Hydroxysteroid dehydrogenase type 1 (17βHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-β-estradiol. 17βHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17βHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17βHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17βHSD1 is a target for identification of lead compounds for structure-based drug design. 17βHSD1 was purified from human placenta. 17βHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 μM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17βHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17βHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
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Gossypol: prototype of inhibitors targeted to dinucleotide Folds.
Current medicinal chemistry, 2000Co-Authors: David L. Vander Jagt, Lorraine M. Deck, Robert E. RoyerAbstract:Gossypol, a disesquiterpene from cottonseed, exhibits multiple biological properties, including male antifertility activity and anticancer activity. Gossypol also inhibits the growth of numerous parasitic organisms and shows antiviral activity against a number of enveloped viruses, including the AIDS virus. Derivatives of gossypol, in which the aldehyde functional groups that contribute to toxicity have been modified, retain or even show enhanced biological activity. Ring substituted 2,3-dihydroxy-1-naphthoic acids, which are structural analogs of gossypol, share with gossypol the ability to complex with dehydrogenases at the dinucleotide Fold (Rossmann Fold) with selectivity, suggesting that gossypol may be considered the prototype of a new class of drugs targeted to dehydrogenases. Most of the biological activities of gossypol and related compounds may result from inhibition of dehydrogenases.
William M. Brown - One of the best experts on this subject based on the ideXlab platform.
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17-beta-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold.
Chemico-biological interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L Vander JagtAbstract:17-beta-Hydroxysteroid dehydrogenase type 1 (17betaHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-beta-estradiol. 17betaHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17betaHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17betaHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17betaHSD1 is a target for identification of lead compounds for structure-based drug design. 17betaHSD1 was purified from human placenta. 17betaHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 microM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17betaHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17betaHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
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17-β-Hydroxysteroid dehydrogenase type 1: computational design of active site inhibitors targeted to the Rossmann Fold
Chemico-Biological Interactions, 2003Co-Authors: William M. Brown, Lorraine M. Deck, Robert E. Royer, Louis E Metzger, Jeremy P. Barlow, Lucy A. Hunsaker, David L. Vander JagtAbstract:Abstract 17-β-Hydroxysteroid dehydrogenase type 1 (17βHSD1), also called estradiol dehydrogenase, catalyzes the NADPH-dependent reduction of the weak estrogen, estrone, into the more potent estrogen, 17-β-estradiol. 17βHSD1 is an attractive drug target in hormone-sensitive breast cancer. Past efforts to develop selective inhibitors of 17βHSD1 have focused on design of substrate analogs. It is challenging to develop steroid analogs that are devoid of any undesired biological activity. 17βHSD1 is a member of the short-chain dehydrogenase/reductase (SDR) superfamily that includes many hydroxysteroid dehydrogenases. Members of the SDR family bind NAD(P)(H) in a motif that is a modified Rossmann Fold. We demonstrated previously that the Rossmann Folds of classical dehydrogenases can be selectively inhibited by derivatives and analogs of the natural product gossypol. In this study, we have addressed the question whether the modified Rossmann Fold in 17βHSD1 is a target for identification of lead compounds for structure-based drug design. 17βHSD1 was purified from human placenta. 17βHSD1 is inhibited by derivatives of gossypol with dissociation constants as low as 2 μM. Inhibition is competitive with the binding of cofactor. Molecular modeling studies using the published coordinates of human 17βHSD1 suggest that these inhibitors occupy the modified Rossmann Fold at the nicotinamide end of the dinucleotide-binding site, extending towards the substrate site. A computational approach was used to design potential new inhibitors of 17βHSD1. The results suggest not only that derivatives of gossypol represent attractive lead compounds for structure-based drug design but also suggest that appropriate incorporation of a substrate analog into the design of these Rossmann Fold inhibitors may provide pan-active site inhibitors that span the cofactor and substrate site, potentially offering specificity and increased potency.
