Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase
Human DHRS6 is a previously uncharacterized member of the short chain dehydrogenases/reductase family and displays significant homologies to bacterial hydroxybutyrate dehydrogenases. Substrate screening reveals sole NAD(+)-dependent conversion of (R)-hydroxybutyrate to acetoacetate with K(m) values...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
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American Society for Biochemistry and Molecular Biology
2006
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| Subjects: |
| _version_ | 1797058146654486528 |
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| author | Guo, K Lukacik, P Papagrigoriou, E Meier, M Lee, W Adamski, J Oppermann, U |
| author_facet | Guo, K Lukacik, P Papagrigoriou, E Meier, M Lee, W Adamski, J Oppermann, U |
| author_sort | Guo, K |
| collection | OXFORD |
| description | Human DHRS6 is a previously uncharacterized member of the short chain dehydrogenases/reductase family and displays significant homologies to bacterial hydroxybutyrate dehydrogenases. Substrate screening reveals sole NAD(+)-dependent conversion of (R)-hydroxybutyrate to acetoacetate with K(m) values of about 10 mm, consistent with plasma levels of circulating ketone bodies in situations of starvation or ketoacidosis. The structure of human DHRS6 was determined at a resolution of 1.8 A in complex with NAD(H) and reveals a tetrameric organization with a short chain dehydrogenases/reductase-typical folding pattern. A highly conserved triad of Arg residues ("triple R" motif consisting of Arg(144), Arg(188), and Arg(205)) was found to bind a sulfate molecule at the active site. Docking analysis of R-beta-hydroxybutyrate into the active site reveals an experimentally consistent model of substrate carboxylate binding and catalytically competent orientation. GFP reporter gene analysis reveals a cytosolic localization upon transfection into mammalian cells. These data establish DHRS6 as a novel, cytosolic type 2 (R)-hydroxybutyrate dehydrogenase, distinct from its well characterized mitochondrial type 1 counterpart. The properties determined for DHRS6 suggest a possible physiological role in cytosolic ketone body utilization, either as a secondary system for energy supply in starvation or to generate precursors for lipid and sterol synthesis. |
| first_indexed | 2024-03-06T19:46:26Z |
| format | Journal article |
| id | oxford-uuid:22735ae1-f9f9-4713-b77e-8ec11769b37d |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:46:26Z |
| publishDate | 2006 |
| publisher | American Society for Biochemistry and Molecular Biology |
| record_format | dspace |
| spelling | oxford-uuid:22735ae1-f9f9-4713-b77e-8ec11769b37d2022-03-26T11:38:55ZCharacterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenaseJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:22735ae1-f9f9-4713-b77e-8ec11769b37dOxidoreductasesArginineDose-Response Relationship, DrugExonsgeneticsLipidsenzymologyHydrogen-Ion ConcentrationGreen Fluorescent ProteinsHela CellsHydroxybutyrate DehydrogenaseCrystallography, X-RayProtein Structure, TertiaryCloning, MolecularAmino Acid MotifsCytosolmetabolismKineticsSulfatesSequence Homology, Amino AcidProtein ConformationAmino Acid SequencePhylogenyMolecular Sequence DataMitochondriaBinding SitesProtein FoldingModels, MolecularchemistrySubstrate SpecificityEnglishStructural Genomics ConsortiumAmerican Society for Biochemistry and Molecular Biology2006Guo, KLukacik, PPapagrigoriou, EMeier, MLee, WAdamski, JOppermann, UHuman DHRS6 is a previously uncharacterized member of the short chain dehydrogenases/reductase family and displays significant homologies to bacterial hydroxybutyrate dehydrogenases. Substrate screening reveals sole NAD(+)-dependent conversion of (R)-hydroxybutyrate to acetoacetate with K(m) values of about 10 mm, consistent with plasma levels of circulating ketone bodies in situations of starvation or ketoacidosis. The structure of human DHRS6 was determined at a resolution of 1.8 A in complex with NAD(H) and reveals a tetrameric organization with a short chain dehydrogenases/reductase-typical folding pattern. A highly conserved triad of Arg residues ("triple R" motif consisting of Arg(144), Arg(188), and Arg(205)) was found to bind a sulfate molecule at the active site. Docking analysis of R-beta-hydroxybutyrate into the active site reveals an experimentally consistent model of substrate carboxylate binding and catalytically competent orientation. GFP reporter gene analysis reveals a cytosolic localization upon transfection into mammalian cells. These data establish DHRS6 as a novel, cytosolic type 2 (R)-hydroxybutyrate dehydrogenase, distinct from its well characterized mitochondrial type 1 counterpart. The properties determined for DHRS6 suggest a possible physiological role in cytosolic ketone body utilization, either as a secondary system for energy supply in starvation or to generate precursors for lipid and sterol synthesis. |
| spellingShingle | Oxidoreductases Arginine Dose-Response Relationship, Drug Exons genetics Lipids enzymology Hydrogen-Ion Concentration Green Fluorescent Proteins Hela Cells Hydroxybutyrate Dehydrogenase Crystallography, X-Ray Protein Structure, Tertiary Cloning, Molecular Amino Acid Motifs Cytosol metabolism Kinetics Sulfates Sequence Homology, Amino Acid Protein Conformation Amino Acid Sequence Phylogeny Molecular Sequence Data Mitochondria Binding Sites Protein Folding Models, Molecular chemistry Substrate Specificity Guo, K Lukacik, P Papagrigoriou, E Meier, M Lee, W Adamski, J Oppermann, U Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase |
| title | Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase |
| title_full | Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase |
| title_fullStr | Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase |
| title_full_unstemmed | Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase |
| title_short | Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase |
| title_sort | characterization of human dhrs6 an orphan short chain dehydrogenase reductase enzyme a novel cytosolic type 2 r beta hydroxybutyrate dehydrogenase |
| topic | Oxidoreductases Arginine Dose-Response Relationship, Drug Exons genetics Lipids enzymology Hydrogen-Ion Concentration Green Fluorescent Proteins Hela Cells Hydroxybutyrate Dehydrogenase Crystallography, X-Ray Protein Structure, Tertiary Cloning, Molecular Amino Acid Motifs Cytosol metabolism Kinetics Sulfates Sequence Homology, Amino Acid Protein Conformation Amino Acid Sequence Phylogeny Molecular Sequence Data Mitochondria Binding Sites Protein Folding Models, Molecular chemistry Substrate Specificity |
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