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ヨコヤマ ケン
YOKOYAMA KEN
横山 謙 所属 京都産業大学 生命科学部 先端生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2015/09/01 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | GraDeR: Membrane Protein Complex Preparation for Single-Particle Cryo-EM. |
| 執筆形態 | その他 |
| 掲載誌名 | Structure (London, England : 1993) |
| 掲載区分 | 国外 |
| 巻・号・頁 | 23(9),pp.1769-1775 |
| 著者・共著者 | Florian Hauer,Christoph Gerle,Niels Fischer,Atsunori Oshima,Kyoko Shinzawa-Itoh,Satoru Shimada,Ken Yokoyama,Yoshinori Fujiyoshi,Holger Stark |
| 概要 | We developed a method, named GraDeR, which substantially improves the preparation of membrane protein complexes for structure determination by single-particle cryo-electron microscopy (cryo-EM). In GraDeR, glycerol gradient centrifugation is used for the mild removal of free detergent monomers and micelles from lauryl maltose-neopentyl glycol detergent stabilized membrane complexes, resulting in monodisperse and stable complexes to which standard processes for water-soluble complexes can be applied. We demonstrate the applicability of the method on three different membrane complexes, including the mammalian FoF1 ATP synthase. For this highly dynamic and fragile rotary motor, we show that GraDeR allows visualizing the asymmetry of the F1 domain, which matches the ground state structure of the isolated domain. Therefore, the present cryo-EM structure of FoF1 ATP synthase provides direct structural evidence for Boyer's binding change mechanism in the context of the intact enzyme. |
| DOI | 10.1016/j.str.2015.06.029 |
| ISSN | 0969-2126 |
| PMID | 26278176 |