論文 - 詳細
| RRC ID | 55157 |
|---|---|
| 著者 | Yoneda K, Sakuraba H, Araki T, Ohshima T. |
| タイトル | Crystal structure of the NADP+ and tartrate-bound complex of L-serine 3-dehydrogenase from the hyperthermophilic archaeon Pyrobaculum calidifontis. |
| ジャーナル | Extremophiles |
| Abstract |
A gene encoding L-serine dehydrogenase (L-SerDH) that exhibits extremely low sequence identity to the Agrobacterium tumefaciens L-SerDH was identified in the hyperthermophilic archaeon Pyrobaculum calidifontis. The predicted amino acid sequence showed 36% identity with that of Pseudomonas aeruginosa L-SerDH, suggesting that P. calidifontis L-SerDH is a novel type of L-SerDH, like Ps. aeruginosa L-SerDH. The overexpressed enzyme appears to be the most thermostable L-SerDH described to date, and no loss of activity was observed by incubation for 30 min at temperatures up to 100 °C. The enzyme showed substantial reactivity towards D-serine, in addition to L-serine. Two different crystal structures of P. calidifontis L-SerDH were determined using the Se-MAD and MR method: the structure in complex with NADP+/sulfate ion at 1.18 Å and the structure in complex with NADP+/L-tartrate (substrate analog) at 1.57 Å. The fold of the catalytic domain showed similarity with that of Ps. aeruginosa L-SerDH. However, the active site structure significantly differed between the two enzymes. Based on the structure of the tartrate, L- and D-serine and 3-hydroxypropionate molecules were modeled into the active site and the substrate binding modes were estimated. A structural comparison suggests that the wide cavity at the substrate binding site is likely responsible for the high reactivity of the enzyme toward both L- and D-serine enantiomers. This is the first description of the structure of the novel type of L-SerDH with bound NADP+ and substrate analog, and it provides new insight into the substrate binding mechanism of L-SerDH. The results obtained here may be very informative for the creation of L- or D-serine-specific SerDH by protein engineering. |
| 巻・号 | 22(3) |
| ページ | 395-405 |
| 公開日 | 2018-5-1 |
| DOI | 10.1007/s00792-018-1004-0 |
| PII | 10.1007/s00792-018-1004-0 |
| PMID | 29353380 |
| MeSH | Alcohol Oxidoreductases / chemistry* Alcohol Oxidoreductases / metabolism Archaeal Proteins / chemistry* Archaeal Proteins / metabolism Catalytic Domain Crystallography, X-Ray Enzyme Stability Hot Temperature Molecular Docking Simulation* NADP / chemistry NADP / metabolism Protein Binding Pyrobaculum / enzymology* Serine / chemistry Serine / metabolism Substrate Specificity Tartrates / chemistry Tartrates / metabolism |
| IF | 2.046 |
| 引用数 | 1 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 一般微生物 | JCM 11548 |