The 1.25Acrystal structure of sepiapterin reductase reveals its binding mode to pterins and brain neurotransmitters

Auerbach, Günter

doi:10.1093/emboj/16.24.7219

Cited by 87 publications

(80 citation statements)

References 63 publications

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“…3 and illustrate the essential nature of these residues for correct coenzyme positioning and binding (Tables I and II). No direct interactions of Thr-12, Asn-86, and Asn-87 with the coenzyme are observed in our model and in most other coenzyme complexes (9,10,(17)(18)(19)). An exception is the recently determined porcine carbonyl reductase, where backbone carbonyl interactions of Asn-89 to the 3Ј-OH of the nicotinamide ribose are found (20).…”

Section: Crystallographic Analysis Of 3␤/17␤-hsd and Comparison With mentioning

confidence: 46%

Critical Residues for Structure and Catalysis in Short-chain Dehydrogenases/Reductases

Filling¹,

Berndt²,

Benach³

et al. 2002

Journal of Biological Chemistry

505

537

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Short-chain dehydrogenases/reductases form a large, evolutionarily old family of NAD(P)(H)-dependent enzymes with over 60 genes found in the human genome. Despite low levels of sequence identity (often 10 -30%), the three-dimensional structures display a highly similar ␣/␤ folding pattern. We have analyzed the role of several conserved residues regarding folding, stability, steady-state kinetics, and coenzyme binding using bacterial 3␤/17␤-hydroxysteroid dehydrogenase and selected mutants. Structure determination of the wildtype enzyme at 1.2-Å resolution by x-ray crystallography and docking analysis was used to interpret the biochemical data. Enzyme kinetic data from mutagenetic replacements emphasize the critical role of residues Thr-12, Asp-60, Asn-86, Asn-87, and Ala-88 in coenzyme binding and catalysis. The data also demonstrate essential interactions of Asn-111 with active site residues. A general role of its side chain interactions for maintenance of the active site configuration to build up a proton relay system is proposed. This extends the previously recognized catalytic triad of Ser-Tyr-Lys residues to form a tetrad of Asn-Ser-Tyr-Lys in the majority of characterized short-chain dehydrogenases/reductase enzymes.

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Section: Crystallographic Analysis Of 3␤/17␤-hsd and Comparison With mentioning

confidence: 46%

Critical Residues for Structure and Catalysis in Short-chain Dehydrogenases/Reductases

Filling¹,

Berndt²,

Benach³

et al. 2002

Journal of Biological Chemistry

505

537

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“…A sample of the same compound, as well as 7-oxo-6-carboxypterin and 7-oxobiopterin isolated from carp skin were donated by Dr. M. Nakagoshi (Kitasato). Construction of the plasmids used for expression of murine recombinant GTP cyclohydrolase I and for murine sepiapterin reductase followed the procedures described earlier (11,32). Recombinant 6-pyruvoyl-H 4 pterin synthase was a gift from Dr. A. Bacher (Mü nchen).…”

Section: Methodsmentioning

confidence: 99%

“…For preparation of 6-pyruvoyl-H 4 pterin as a potential substrate for the xanthine oxidoreductase variant, H 2 neopterin triphosphate was first generated by incubation of GTP with recombinant murine GTP cyclohydrolase I, and EDTA was quenched by addition of Mg 2ϩ as described above; the reaction mixture was incubated with recombinant 6-pyruvoyl-H 4 pterin synthase for 20 min and then immediately used. Due to the endogenous sepiapterin reductase activity of zebrafish homogenates and the additional function of this enzyme as an isomerase (11,37), the intermediates 6-lactoyl-H 4 pterin and 6-(1Ј-hydroxy-2Ј-oxopropyl)-H 4 pterin are transiently formed also, so that these could also be considered as possible precursors for 7-oxobiopterin formation. The assay for xanthine oxidoreductase activity used the conversion of pterin to isoxanthopterin, which was then separated by HPLC (solvent system IV; see legend of Table I) and fluorometrically detected (340-nm excitation/410-nm emission) (38,39).…”

Section: Methodsmentioning

confidence: 99%

Development of the Pteridine Pathway in the Zebrafish,Danio rerio

Ziegler¹,

McDonald²,

Heßlinger³

et al. 2000

Journal of Biological Chemistry

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In the zebrafish, the peripheral neurons and the pigment cells are derived from the neural crest and share the pteridine pathway, which leads either to the cofactor tetrahydrobiopterin or to xanthophore pigments. The components of the pteridine pattern were identified as tetrahydrobiopterin, sepiapterin, 7-oxobiopterin, isoxanthopterin, and 2,4,7-trioxopteridine. The expression of GTP cyclohydrolase I activity during the first 24-h postfertilization, followed by 6-pyruvoyl-5,6,7,8-tetrahydropterin synthase and sepiapterin reductase, suggest an early supply of tetrahydrobiopterin for neurotransmitter synthesis in the neurons and for tyrosine supply in the melanophores. At 48-h postfertilization, sepiapterin formation branches off the de novo pathway of tetrahydrobiopterin synthesis. Sepiapterin, via 7,8-dihydrobiopterin and biopterin, serves as a precursor for the formation of 7-oxobiopterin, which may be further catabolized to isoxanthopterin and 2,4,7-trioxopteridine. Neither 7,8-dihydrobiopterin nor biopterin is a substrate for xanthine oxidoreductase. In contrast, both of these compounds are oxidized at C-7 by a xanthine oxidase variant form, which is inactivated by KCN, but is insensitive to allopurinol. The oxidase and the dehydrogenase form of xanthine oxidoreductase as well as the xanthine oxidase variant have specific developmental patterns. It follows that GTP cyclohydrolase I, the formation of sepiapterin, and the xanthine oxidoreductase family control the pteridine pathway in the zebrafish.

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“…Crystallographic studies with SPR complexed with sulfasalazine, sulfapyridine, or sulfathiazole and other inhibitors have provided insight into the mechanism underlying inhibition of sepiapterin reduction and redox cycling (Auerbach et al, 1997;Haruki et al, 2013). A highly conserved sequence motif in the catalytic site of the human enzyme is highlighted by a critical triad pocket composed of Ser157, Tyr170, and Lys174 (Ser158, Tyr171 and Lys175 in the mouse enzyme), which is important in stabilizing protein structure, maintaining substrate/cofactor proximity, and proton transfer (Auerbach et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

Sulfa Drugs Inhibit Sepiapterin Reduction and Chemical Redox Cycling by Sepiapterin Reductase

Yang

Jan

Mishin

et al. 2014

J Pharmacol Exp Ther

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Sepiapterin reductase (SPR) catalyzes the reduction of sepiapterin to dihydrobiopterin (BH2), the precursor for tetrahydrobiopterin (BH4), a cofactor critical for nitric oxide biosynthesis and alkylglycerol and aromatic amino acid metabolism. SPR also mediates chemical redox cycling, catalyzing one-electron reduction of redox-active chemicals, including quinones and bipyridinium herbicides (e.g., menadione, 9,10-phenanthrenequinone, and diquat); rapid reaction of the reduced radicals with molecular oxygen generates reactive oxygen species (ROS). Using recombinant human SPR, sulfonamide-and sulfonylurea-based sulfa drugs were found to be potent noncompetitive inhibitors of both sepiapterin reduction and redox cycling. The most potent inhibitors of sepiapterin reduction (IC 50 s 5 31-180 nM) were sulfasalazine, sulfathiazole, sulfapyridine, sulfamethoxazole, and chlorpropamide. Higher concentrations of the sulfa drugs (IC 50 s 5 0.37-19.4 mM) were required to inhibit redox cycling, presumably because of distinct mechanisms of sepiapterin reduction and redox cycling. In PC12 cells, which generate catecholamine and monoamine neurotransmitters via BH4-dependent amino acid hydroxylases, sulfa drugs inhibited both BH2/BH4 biosynthesis and redox cycling mediated by SPR. Inhibition of BH2/BH4 resulted in decreased production of dopamine and dopamine metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, and 5-hydroxytryptamine. Sulfathiazole (200 mM) markedly suppressed neurotransmitter production, an effect reversed by BH4. These data suggest that SPR and BH4-dependent enzymes, are "off-targets" of sulfa drugs, which may underlie their untoward effects. The ability of the sulfa drugs to inhibit redox cycling may ameliorate ROS-mediated toxicity generated by redox active drugs and chemicals, contributing to their anti-inflammatory activity.

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The 1.25Acrystal structure of sepiapterin reductase reveals its binding mode to pterins and brain neurotransmitters

Cited by 87 publications

References 63 publications

Critical Residues for Structure and Catalysis in Short-chain Dehydrogenases/Reductases

Critical Residues for Structure and Catalysis in Short-chain Dehydrogenases/Reductases

Development of the Pteridine Pathway in the Zebrafish,Danio rerio

Sulfa Drugs Inhibit Sepiapterin Reduction and Chemical Redox Cycling by Sepiapterin Reductase

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