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-Structure paper
タイトル | A 'Build and Retrieve' methodology to simultaneously solve cryo-EM structures of membrane proteins. |
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ジャーナル・号・ページ | Nat Methods, Vol. 18, Issue 1, Page 69-75, Year 2021 |
掲載日 | 2021年1月6日 |
著者 | Chih-Chia Su / Meinan Lyu / Christopher E Morgan / Jani Reddy Bolla / Carol V Robinson / Edward W Yu / |
PubMed 要旨 | Single-particle cryo-electron microscopy (cryo-EM) has become a powerful technique in the field of structural biology. However, the inability to reliably produce pure, homogeneous membrane protein ...Single-particle cryo-electron microscopy (cryo-EM) has become a powerful technique in the field of structural biology. However, the inability to reliably produce pure, homogeneous membrane protein samples hampers the progress of their structural determination. Here, we develop a bottom-up iterative method, Build and Retrieve (BaR), that enables the identification and determination of cryo-EM structures of a variety of inner and outer membrane proteins, including membrane protein complexes of different sizes and dimensions, from a heterogeneous, impure protein sample. We also use the BaR methodology to elucidate structural information from Escherichia coli K12 crude membrane and raw lysate. The findings demonstrate that it is possible to solve high-resolution structures of a number of relatively small (<100 kDa) and less abundant (<10%) unidentified membrane proteins within a single, heterogeneous sample. Importantly, these results highlight the potential of cryo-EM for systems structural proteomics. |
リンク | Nat Methods / PubMed:33408407 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.38 - 3.6 Å |
構造データ | EMDB-21897, PDB-6wti: EMDB-21900, PDB-6wtz: EMDB-21901, PDB-6wu0: EMDB-21906, PDB-6wu6: EMDB-22528, PDB-7jz2: EMDB-22529, PDB-7jz3: EMDB-22530, PDB-7jz6: EMDB-22531, PDB-7jzh: |
化合物 | ChemComp-HEM: ChemComp-HEO: ChemComp-CU: ChemComp-UQ8: ChemComp-3PE: ChemComp-U9V: ChemComp-HOH: ChemComp-FES: ChemComp-SF4: ChemComp-F3S: ChemComp-FAD: ChemComp-NA: ChemComp-UQ2: |
由来 |
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キーワード | OXIDOREDUCTASE / ubiquinol oxidase cytochrome bo3 / membrane protein / outer membrane porin / omp / ompf / PROTEIN TRANSPORT / CryoEM structure / hopanoid transporter HpnN / ELECTRON TRANSPORT / Complex / succinate-coenzyme Q reductase / ELECTRON TRANSPORT/OXIDOREDUCTASE / Succinate: quinone oxidoreductase / SdhA / sdhB / SdhC / SdhD / ELECTRON TRANSPORT-OXIDOREDUCTASE complex / TRANSPORT PROTEIN / Osmoporin OmpC / catalase-peroxidase / LYASE / Glutamate decarboxylase beta |