Critical residues for structure and catalysis in short-chain dehydrogenases/reductases.
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 alpha/beta folding pattern. We...
| Hauptverfasser: | , , , , , , , , |
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| Format: | Journal article |
| Sprache: | English |
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2002
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| _version_ | 1826294365985701888 |
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| author | Filling, C Berndt, K Benach, J Knapp, S Prozorovski, T Nordling, E Ladenstein, R Jörnvall, H Oppermann, U |
| author_facet | Filling, C Berndt, K Benach, J Knapp, S Prozorovski, T Nordling, E Ladenstein, R Jörnvall, H Oppermann, U |
| author_sort | Filling, C |
| collection | OXFORD |
| description | 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 alpha/beta folding pattern. We have analyzed the role of several conserved residues regarding folding, stability, steady-state kinetics, and coenzyme binding using bacterial 3beta/17beta-hydroxysteroid dehydrogenase and selected mutants. Structure determination of the wild-type enzyme at 1.2-A 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. |
| first_indexed | 2024-03-07T03:44:31Z |
| format | Journal article |
| id | oxford-uuid:bf01b20c-74f8-4f17-87e5-f3381c9db8db |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:44:31Z |
| publishDate | 2002 |
| record_format | dspace |
| spelling | oxford-uuid:bf01b20c-74f8-4f17-87e5-f3381c9db8db2022-03-27T05:44:15ZCritical residues for structure and catalysis in short-chain dehydrogenases/reductases.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bf01b20c-74f8-4f17-87e5-f3381c9db8dbEnglishSymplectic Elements at Oxford2002Filling, CBerndt, KBenach, JKnapp, SProzorovski, TNordling, ELadenstein, RJörnvall, HOppermann, UShort-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 alpha/beta folding pattern. We have analyzed the role of several conserved residues regarding folding, stability, steady-state kinetics, and coenzyme binding using bacterial 3beta/17beta-hydroxysteroid dehydrogenase and selected mutants. Structure determination of the wild-type enzyme at 1.2-A 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. |
| spellingShingle | Filling, C Berndt, K Benach, J Knapp, S Prozorovski, T Nordling, E Ladenstein, R Jörnvall, H Oppermann, U Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. |
| title | Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. |
| title_full | Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. |
| title_fullStr | Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. |
| title_full_unstemmed | Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. |
| title_short | Critical residues for structure and catalysis in short-chain dehydrogenases/reductases. |
| title_sort | critical residues for structure and catalysis in short chain dehydrogenases reductases |
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