The Crystal Structure of 3α-Hydroxysteroid Dehydrogenase/Carbonyl Reductase from Comamonas testosteroni Shows a Novel Oligomerization Pattern within the Short Chain Dehydrogenase/Reductase Family

Grimm, Clemens; Maser, Edmund; Möbus, Eric; Klebe, G.; Reuter, Klaus; Ficner, Ralf

doi:10.1074/jbc.m007559200

Cited by 96 publications

(98 citation statements)

References 32 publications

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“…For all convergently evolved RDH enzymes from the SDR superfamily, only one conformational orientation of dinucleotides within the binding site has been observed. This is consistent with the structural conservation and rigidity of the Rossman fold (33)(34)(35)(36)(37). However, the binding of hydrophobic substrates occurs in less conserved loop regions and can take place by projecting the aldehyde group in re-or si-face orientation toward the nucleotide (24).…”

Section: Dual-substrate Specificity Dual Responsibility; Mechanisticsupporting

confidence: 62%

Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina

Haeseleer¹,

Jang²,

Imanishi³

et al. 2002

Journal of Biological Chemistry

184

227

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Retinoids are chromophores involved in vision, transcriptional regulation, and cellular differentiation. Members of the short chain alcohol dehydrogenase/reductase superfamily catalyze the transformation of retinol to retinal. Here, we describe the identification and properties of three enzymes from a novel subfamily of four retinol dehydrogenases (RDH11-14) that display dualsubstrate specificity, uniquely metabolizing all-trans-and cis-retinols with C 15 pro-R specificity. RDH11-14 could be involved in the first step of all-trans-and 9-cis-retinoic acid production in many tissues. RDH11-14 fill the gap in our understanding of 11-cis-retinal and all-trans-retinal transformations in photoreceptor (RDH12) and retinal pigment epithelial cells (RDH11). The dualsubstrate specificity of RDH11 explains the minor phenotype associated with mutations in 11-cisretinol dehydrogenase (RDH5) causing fundus albipunctatus in humans and engineered mice lacking RDH5. Furthermore, photoreceptor RDH12 could be involved in the production of 11-cis-retinal from 11-cis-retinol during regeneration of the cone visual pigments. These newly identified enzymes add new elements to important retinoid metabolic pathways that have not been explained by previous genetic and biochemical studies.Retinoids are indispensable light-sensitive elements of vision and also serve as essential modulators of cellular differentiation and proliferation in diverse cell types, including those comprising the epithelium and immune system. Retinoids modulate the growth of both normal and malignant cells through their binding to retinoid receptors. All-trans-retinoic acid signals through specific interactions with the nuclear retinoic acid receptors, whereas its isomer, 9-cis-retinoic acid, is a high affinity ligand of retinoic acid receptors and retinoid X receptors. In the retina, light-dependent photoisomerization of 11-cis-retinylidene to the all-transretinylidene moiety of rod and cone photoreceptors is a key reaction that triggers visual * This research was supported by National Institutes of Health Grants EY08061, CA75173, and DK59125, a grant from Research to Prevent Blindness, Inc. (to the Department of Ophthalmology, University of Washington), and by the E. K. Bishop Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. S The on-line version of this article (available at http://www.jbc.org) contains Supplemental Figs. 1 and 2 and Tables 1-3. § These authors contributed equally to this work. § § Recipient of a Research to Prevent Blindness Senior Investigator Award. To whom correspondence should be addressed: Dept. of Ophthalmology, University of Washington, Seattle,; E-mail: palczews@u.washington.edu. Transformations of retinoids occur mostly through enzymatic or photochemical reactions, although they readily isomerize non-enzymatically to thermodynamic equilibriu...

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Section: Dual-substrate Specificity Dual Responsibility; Mechanisticsupporting

confidence: 62%

Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina

Haeseleer¹,

Jang²,

Imanishi³

et al. 2002

Journal of Biological Chemistry

184

227

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“…The structures of the binary complex with NAD ϩ cofactor and apoenzyme of 3␣-HSD/CR have been solved (3,17). In the binary complex, the NAD ϩ cofactor is bound at the C-terminal ends of the ␤-strands in the 3␣-HSD/CR from C. testosterone.…”

mentioning

confidence: 99%

“…Kinetically, the 3␣-HSD/CR-catalyzed reaction shows an ordered bi bi mechanism with the pyrimidine nucleotide being the first substrate to bind and the last product to be released (18). The chemical mechanism for the 3␣-HSD/ CR-catalyzed reaction is based on sequence comparisons and structure analyses in the SDR family (17). A triad of Ser-114, Tyr-155, and Lys-159 in 3␣-HSD/CR is conserved within the SDR family.…”

mentioning

confidence: 99%

Mechanistic Roles of Ser-114, Tyr-155, and Lys-159 in 3α-Hydroxysteroid Dehydrogenase/Carbonyl Reductase from Comamonas testosteroni

Hwang

Chang

Hsu

et al. 2005

Journal of Biological Chemistry

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3␣-Hydroxysteroid dehydrogenase/carbonyl reductase (3␣-HSD/CR) from؊5 times of wild-type activity at pH 10.5. The activity decreases as the pH lowers, which indicates that a functional group with an apparent pK a of 7.2 is involved in the general base catalysis for wild-type 3␣-HSD/CR. The pK a shift to 9.1 for mutant K159A suggests the role of Lys-159 is to lower the pK a of the residues involved in the general base catalysis. Because pH dependence is observed for both S114A and Y155F mutants and pH independence is observed in S114A/Y155F, Tyr-155 may be important as a general base catalysis in the wild-type, whereas Ser-114 may act as a general base on mutant Y155F to catalyze the reaction.

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“…To further test and verify the steroid biodegradability by strain H5, 3 H labeled testosterone and estradiol were used in the present study. C. testosteroni ATCC1196 was employed as positive control to confirm that the procedure involved is competent in proving the steroid biodegradability by strain H5 [20][21][22][23][31][32][33][34][35][36][37][38][39][40].…”

Section: Steroid Biodegradability By Marine Strain H5mentioning

confidence: 99%

Steroid degradation and two steroid-inducible enzymes in the marine bacterium H5

Sang

Guan²,

Maser³

2011

Chemico-Biological Interactions

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a b s t r a c tNatural and synthetic steroid hormones excreted into the environment are potentially threatening the population dynamics of all kinds of animals and public health. We have previously isolated a steroid degrading bacterial strain (H5) from the Baltic Sea, at Kiel, Germany. 16S-rRNA analysis showed that bacterial strain H5 belongs to the genus Vibrio, family Vibrionaceae and class Gamma-Proteobacteria. Bacterial strain H5 can degrade steroids such as testosterone and estrogens, which was shown in this study by determining the 3 H labeled steroid retaining in the bacterial H5 culture medium at incubation times of 5 h and 20 h. Since 3␣-hydroxysteroid dehydrogenase/carbonyl reductase (3␣-HSD/CR) is a key enzyme in adaptive steroid degradation in Comamonas testosteroni (C. testosteroni), in previous investigations, a meta-genomic system with the 3␣-HSD/CR gene as a positive control was established. By this meta-genomic system, two estradiol inducible genes coding 3-ketosteroid-delta-1-dehydrogenase and carboxylesterase, respectively, which are involved in steroid degradation, were found in marine strain H5. In the present work, the 3-ketosteroid-delta-1-dehydrogenase and carboxylesterase genes were subcloned into plasmids pET38-12 and pET24-17, respectively. Overexpression in Escherichia coli (E. coli) strain BL21(DE3)pLysS cells resulted in corresponding proteins with an N-terminal His-tag sequence. After induction with isopropyl-␤-D-thiogalactoside, 3-ketosteroid-delta-1-dehydrogenase and carboxylesterase were purified in one step using nickel-chelate chromatography. After protein determination, 3-ketosteroid-delta-1-dehydrogenase (0.48 mg/ml) and carboxylesterase (1.28 mg/ml) were used to prepare antibodies to determine steroid binding specificity in future research. In summary, we have shown that the marine strain H5 could metabolize steroids; have isolated two estradiol inducible genes from strain H5 chromosomal DNA, and purified the corresponding proteins for further research. The exact characterization and systematic classification of the marine steroid degrading bacterial strain H5 is envisaged. The strain might be used for the bioremediation of steroid contaminations in seawater.

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The Crystal Structure of 3α-Hydroxysteroid Dehydrogenase/Carbonyl Reductase from Comamonas testosteroni Shows a Novel Oligomerization Pattern within the Short Chain Dehydrogenase/Reductase Family

Cited by 96 publications

References 32 publications

Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina

Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina

Mechanistic Roles of Ser-114, Tyr-155, and Lys-159 in 3α-Hydroxysteroid Dehydrogenase/Carbonyl Reductase from Comamonas testosteroni

Steroid degradation and two steroid-inducible enzymes in the marine bacterium H5

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