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-Structure paper
タイトル | Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis. |
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ジャーナル・号・ページ | Commun Biol, Vol. 4, Issue 1, Page 1337, Year 2021 |
掲載日 | 2021年11月25日 |
著者 | Anna J Higgins / Alex J Flynn / Anaïs Marconnet / Laura J Musgrove / Vincent L G Postis / Jonathan D Lippiat / Chun-Wa Chung / Tom Ceska / Manuela Zoonens / Frank Sobott / Stephen P Muench / |
PubMed 要旨 | Membrane proteins are essential for cellular growth, signalling and homeostasis, making up a large proportion of therapeutic targets. However, the necessity for a solubilising agent to extract them ...Membrane proteins are essential for cellular growth, signalling and homeostasis, making up a large proportion of therapeutic targets. However, the necessity for a solubilising agent to extract them from the membrane creates challenges in their structural and functional study. Although amphipols have been very effective for single-particle electron cryo-microscopy (cryoEM) and mass spectrometry, they rely on initial detergent extraction before exchange into the amphipol environment. Therefore, circumventing this pre-requirement would be a big advantage. Here we use an alternative type of amphipol: a cycloalkane-modified amphiphile polymer (CyclAPol) to extract Escherichia coli AcrB directly from the membrane and demonstrate that the protein can be isolated in a one-step purification with the resultant cryoEM structure achieving 3.2 Å resolution. Together this work shows that cycloalkane amphipols provide a powerful approach for the study of membrane proteins, allowing native extraction and high-resolution structure determination by cryoEM. |
リンク | Commun Biol / PubMed:34824357 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.2 Å |
構造データ | EMDB-12043, PDB-7b5p: |
由来 |
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キーワード | MEMBRANE PROTEIN / drug exporter / amphipol |