Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold.
Short-chain dehydrogenases/reductases (SDR) constitute a large family of enzymes found in all forms of life. Despite a low level of sequence identity, the three-dimensional structures determined display a nearly superimposable alpha/beta folding pattern. We identified a conserved asparagine residue...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2001
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| _version_ | 1797098753051590656 |
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| author | Filling, C Nordling, E Benach, J Berndt, K Ladenstein, R Jörnvall, H Oppermann, U |
| author_facet | Filling, C Nordling, E Benach, J Berndt, K Ladenstein, R Jörnvall, H Oppermann, U |
| author_sort | Filling, C |
| collection | OXFORD |
| description | Short-chain dehydrogenases/reductases (SDR) constitute a large family of enzymes found in all forms of life. Despite a low level of sequence identity, the three-dimensional structures determined display a nearly superimposable alpha/beta folding pattern. We identified a conserved asparagine residue located within strand betaF and analyzed its role in the short-chain dehydrogenase/reductase architecture. Mutagenetic replacement of Asn179 by Ala in bacterial 3beta/17beta-hydroxysteroid dehydrogenase yields a folded, but enzymatically inactive enzyme, which is significantly more resistant to denaturation by guanidinium hydrochloride. Crystallographic analysis of the wild-type enzyme at 1.2-A resolution reveals a hydrogen bonding network, including a buried and well-ordered water molecule connecting strands betaE to betaF, a common feature found in 16 of 21 known three-dimensional structures of the family. Based on these results, we hypothesize that in mammalian 11beta-hydroxysteroid dehydrogenase the essential Asn-linked glycosylation site, which corresponds to the conserved segment, displays similar structural features and has a central role to maintain the SDR scaffold. |
| first_indexed | 2024-03-07T05:14:09Z |
| format | Journal article |
| id | oxford-uuid:dc90a21c-1b3a-475a-878e-21d34504258b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:14:09Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:dc90a21c-1b3a-475a-878e-21d34504258b2022-03-27T09:18:42ZStructural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:dc90a21c-1b3a-475a-878e-21d34504258bEnglishSymplectic Elements at Oxford2001Filling, CNordling, EBenach, JBerndt, KLadenstein, RJörnvall, HOppermann, UShort-chain dehydrogenases/reductases (SDR) constitute a large family of enzymes found in all forms of life. Despite a low level of sequence identity, the three-dimensional structures determined display a nearly superimposable alpha/beta folding pattern. We identified a conserved asparagine residue located within strand betaF and analyzed its role in the short-chain dehydrogenase/reductase architecture. Mutagenetic replacement of Asn179 by Ala in bacterial 3beta/17beta-hydroxysteroid dehydrogenase yields a folded, but enzymatically inactive enzyme, which is significantly more resistant to denaturation by guanidinium hydrochloride. Crystallographic analysis of the wild-type enzyme at 1.2-A resolution reveals a hydrogen bonding network, including a buried and well-ordered water molecule connecting strands betaE to betaF, a common feature found in 16 of 21 known three-dimensional structures of the family. Based on these results, we hypothesize that in mammalian 11beta-hydroxysteroid dehydrogenase the essential Asn-linked glycosylation site, which corresponds to the conserved segment, displays similar structural features and has a central role to maintain the SDR scaffold. |
| spellingShingle | Filling, C Nordling, E Benach, J Berndt, K Ladenstein, R Jörnvall, H Oppermann, U Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. |
| title | Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. |
| title_full | Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. |
| title_fullStr | Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. |
| title_full_unstemmed | Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. |
| title_short | Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. |
| title_sort | structural role of conserved asn179 in the short chain dehydrogenase reductase scaffold |
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