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-Structure paper
タイトル | Structure of Aquifex aeolicus lumazine synthase by cryo-electron microscopy to 1.42 Å resolution. |
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ジャーナル・号・ページ | IUCrJ, Vol. 11, Issue Pt 5, Page 723-729, Year 2024 |
掲載日 | 2024年9月1日 |
著者 | Christos G Savva / Mohamed A Sobhy / Alfredo De Biasio / Samir M Hamdan / |
PubMed 要旨 | Single-particle cryo-electron microscopy (cryo-EM) has become an essential structural determination technique with recent hardware developments making it possible to reach atomic resolution, at which ...Single-particle cryo-electron microscopy (cryo-EM) has become an essential structural determination technique with recent hardware developments making it possible to reach atomic resolution, at which individual atoms, including hydrogen atoms, can be resolved. In this study, we used the enzyme involved in the penultimate step of riboflavin biosynthesis as a test specimen to benchmark a recently installed microscope and determine if other protein complexes could reach a resolution of 1.5 Å or better, which so far has only been achieved for the iron carrier ferritin. Using state-of-the-art microscope and detector hardware as well as the latest software techniques to overcome microscope and sample limitations, a 1.42 Å map of Aquifex aeolicus lumazine synthase (AaLS) was obtained from a 48 h microscope session. In addition to water molecules and ligands involved in the function of AaLS, we can observe positive density for ∼50% of the hydrogen atoms. A small improvement in the resolution was achieved by Ewald sphere correction which was expected to limit the resolution to ∼1.5 Å for a molecule of this diameter. Our study confirms that other protein complexes can be solved to near-atomic resolution. Future improvements in specimen preparation and protein complex stabilization may allow more flexible macromolecules to reach this level of resolution and should become a priority of study in the field. |
リンク | IUCrJ / PubMed:38965901 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 1.42 Å |
構造データ | EMDB-39478, PDB-8yt4: |
化合物 | ChemComp-PO4: ChemComp-HOH: |
由来 |
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キーワード | BIOSYNTHETIC PROTEIN / Enzyme involved in riboflavin biosynthesis |