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| タイトル | Overcoming Preferred Orientation in Cryo-EM With Ultrasonic Excitation During Vitrification. |
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| ジャーナル・号・ページ | bioRxiv, Year 2025 |
| 掲載日 | 2025年9月17日 |
著者 | Harry M Williams / Wyatt A Curtis / Michal Haubner / Jakub Hruby / Marcel Drabbels / Ulrich J Lorenz / ![]() |
| PubMed 要旨 | Preferred particle orientation remains a frequently encountered issue in cryo-electron microscopy that arises when proteins adsorb to the air-water interface in only a limited number of orientations. ...Preferred particle orientation remains a frequently encountered issue in cryo-electron microscopy that arises when proteins adsorb to the air-water interface in only a limited number of orientations. This can significantly increase the data acquisition time required to reach a desired resolution or even make it impossible to obtain a reconstruction altogether. Here, we show that preferred orientation can be overcome by continuously exciting the sample with ultrasonic waves during vitrification. Our experiments suggest that mechanical oscillations induced in the sample support continuously shake proteins loose form the air-water interface, thereby scrambling their orientations. The simple, physical nature of this mechanism should make it applicable to a wide range of proteins. Since our method can be easily implemented in existing vitrification devices, it should find widespread adoption. |
リンク | bioRxiv / PubMed:41000704 / PubMed Central |
| 手法 | EM (単粒子) |
| 解像度 | 2.7 - 4.3 Å |
| 構造データ | ![]() EMDB-54829: Cryo-EM map of 50S ribosomal subunit following ultrasonic excitation ![]() EMDB-54830: Cryo-EM map of 50S ribosomal subunit jetted control sample ![]() EMDB-54831: Cryo-EM map of influenza hemagglutinin (A/Hong Kong/1/1968, H3N2) following ultrasonic excitation ![]() EMDB-54833: Cryo-EM map of human pentameric C-reactive protein following ultrasonic excitation ![]() EMDB-54834: Cryo-EM map of human pentameric C-reactive protein jetted control sample ![]() EMDB-54835: Cryo-EM map of human decameric C-reactive protein following ultrasonic excitation ![]() EMDB-54836: Cryo-EM map of human decameric C-reactive protein jetted control sample |
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Influenza A virus (A/Hong Kong/01/1968(H3N2)) (A型インフルエンザウイルス)
Homo sapiens (ヒト)