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-Structure paper
タイトル | High-resolution structure determination of sub-100 kDa complexes using conventional cryo-EM. |
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ジャーナル・号・ページ | Nat Commun, Vol. 10, Issue 1, Page 1032, Year 2019 |
掲載日 | 2019年3月4日 |
著者 | Mark A Herzik / Mengyu Wu / Gabriel C Lander / |
PubMed 要旨 | Determining high-resolution structures of biological macromolecules amassing less than 100 kilodaltons (kDa) has been a longstanding goal of the cryo-electron microscopy (cryo-EM) community. While ...Determining high-resolution structures of biological macromolecules amassing less than 100 kilodaltons (kDa) has been a longstanding goal of the cryo-electron microscopy (cryo-EM) community. While the Volta phase plate has enabled visualization of specimens in this size range, this instrumentation is not yet fully automated and can present technical challenges. Here, we show that conventional defocus-based cryo-EM methodologies can be used to determine high-resolution structures of specimens amassing less than 100 kDa using a transmission electron microscope operating at 200 keV coupled with a direct electron detector. Our ~2.7 Å structure of alcohol dehydrogenase (82 kDa) proves that bound ligands can be resolved with high fidelity to enable investigation of drug-target interactions. Our ~2.8 Å and ~3.2 Å structures of methemoglobin demonstrate that distinct conformational states can be identified within a dataset for proteins as small as 64 kDa. Furthermore, we provide the sub-nanometer cryo-EM structure of a sub-50 kDa protein. |
リンク | Nat Commun / PubMed:30833564 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.8 - 6.0 Å |
構造データ | EMDB-0406: Single-particle cryo-EM reconstruction of horse liver alcohol dehydrogenase using 200 keV EMDB-0407: Single-particle cryo-EM reconstruction of human methemoglobin using 200 keV, state 1 EMDB-0408: Single-particle cryo-EM reconstruction of human methemoglobin using 200 keV, state 2 EMDB-0409: |
化合物 | ChemComp-NAD: ChemComp-ZN: ChemComp-HEM: |
由来 |
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キーワード | OXIDOREDUCTASE / dehydrogenase / NADH-binding / homo-2-mer / OXYGEN TRANSPORT / heme-binding / hetero-4-mer / globin |