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ツゲ ヒデアキ
TSUGE HIDEAKI
津下 英明 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 | 2021 |
| 形態種別 | 研究論文 |
| 標題 | Preparation of Clostridium perfringens binary iota-toxin pore complex for structural analysis using cryo-EM. |
| 執筆形態 | その他 |
| 掲載誌名 | Methods in enzymology |
| 掲載区分 | 国外 |
| 巻・号・頁 | 649,pp.125-148 |
| 著者・共著者 | Tomohito Yamada,Hideaki Tsuge |
| 概要 | Iota toxin, a type of A-B toxin produced by Clostridium perfringens, comprises an enzymatic component (Ia) and a membrane-binding component (Ib). The translocation of Ia to the target cell via the pore formed by Ib allows it to function as an ADP-ribosyltransferase that inhibits actin polymerization in the host cell. The structure of Ia-bound Ib-pore has been determined using cryo-electron microscopy (cryo-EM), thereby elucidating the mechanism of the initial Ia translocation; however, open questions regarding Ia translocation still exist. In this chapter, we describe a new method of preparing Ia-bound Ib-pore complex samples for structural analysis at high resolution using cryo-EM. This method is different from previously reported methods for other A-B toxins. Consequently, it produces Ib-pore with two different states with short and long membrane-spanning β-barrel stem. We expect that this method will be useful in functional and structural studies of iota toxin and other binary toxins. |
| DOI | 10.1016/bs.mie.2021.01.032 |
| PMID | 33712185 |