論文 - 詳細
| RRC ID | 52859 |
|---|---|
| 著者 | Iwasaki T, Fukazawa R, Miyajima-Nakano Y, Baldansuren A, Matsushita S, Lin MT, Gennis RB, Hasegawa K, Kumasaka T, Dikanov SA. |
| タイトル | Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. |
| ジャーナル | J Am Chem Soc |
| Abstract |
The electronic structure and geometry of redox-active metal cofactors in proteins are tuned by the pattern of hydrogen bonding with the backbone peptide matrix. In this study we developed a method for selective amino acid labeling of a hyperthermophilic archaeal metalloprotein with engineered Escherichia coli auxotroph strains, and we applied this to resolve the hydrogen bond interactions with the reduced Rieske-type [2Fe-2S] cluster by two-dimensional pulsed electron spin resonance technique. Because deep electron spin-echo envelope modulation of two histidine (14)N(δ) ligands of the cluster decreased non-coordinating (15)N signal intensities via the cross-suppression effect, an inverse labeling strategy was employed in which (14)N amino acid-labeled archaeal Rieske-type ferredoxin samples were examined in an (15)N-protein background. This has directly identified Lys45 N(α) as providing the major pathway for the transfer of unpaired electron spin density from the reduced cluster by a "through-bond" mechanism. All other backbone peptide nitrogens interact more weakly with the reduced cluster. The extension of this approach will allow visualizing the three-dimensional landscape of preferred pathways for the transfer of unpaired spin density from a paramagnetic metal center onto the protein frame, and will discriminate specific interactions by a "through-bond" mechanism from interactions which are "through-space" in various metalloproteins. |
| 巻・号 | 134(48) |
| ページ | 19731-8 |
| 公開日 | 2012-12-5 |
| DOI | 10.1021/ja308049u |
| PMID | 23145461 |
| PMC | PMC3515699 |
| MeSH | Binding Sites Escherichia coli / genetics Ferredoxins / chemistry* Hydrogen Bonding Iron / chemistry* Isotope Labeling Models, Molecular Oxidation-Reduction Pyrodictiaceae / chemistry Substrate Specificity Sulfolobus solfataricus / chemistry Sulfur / chemistry* |
| IF | 14.695 |
| 引用数 | 14 |
| 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 | 1 |
| 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 | |
| 遺伝子材料 | YM138 (RDB15943) MS1 (RDB15944). |