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-Structure paper
タイトル | A cryo-EM grid preparation device for time-resolved structural studies. |
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ジャーナル・号・ページ | IUCrJ, Vol. 6, Issue Pt 6, Page 1024-1031, Year 2019 |
掲載日 | 2019年11月1日 |
著者 | Dimitrios Kontziampasis / David P Klebl / Matthew G Iadanza / Charlotte A Scarff / Florian Kopf / Frank Sobott / Diana C F Monteiro / Martin Trebbin / Stephen P Muench / Howard D White / |
PubMed 要旨 | Structural biology generally provides static snapshots of protein conformations that can provide information on the functional mechanisms of biological systems. Time-resolved structural biology ...Structural biology generally provides static snapshots of protein conformations that can provide information on the functional mechanisms of biological systems. Time-resolved structural biology provides a means to visualize, at near-atomic resolution, the dynamic conformational changes that macromolecules undergo as they function. X-ray free-electron-laser technology has provided a powerful tool to study enzyme mechanisms at atomic resolution, typically in the femtosecond to picosecond timeframe. Complementary to this, recent advances in the resolution obtainable by electron microscopy and the broad range of samples that can be studied make it ideally suited to time-resolved approaches in the microsecond to millisecond timeframe to study large loop and domain motions in biomolecules. Here we describe a cryo-EM grid preparation device that permits rapid mixing, voltage-assisted spraying and vitrification of samples. It is shown that the device produces grids of sufficient ice quality to enable data collection from single grids that results in a sub-4 Å reconstruction. Rapid mixing can be achieved by blot-and-spray or mix-and-spray approaches with a delay of ∼10 ms, providing greater temporal resolution than previously reported mix-and-spray approaches. |
リンク | IUCrJ / PubMed:31709058 / PubMed Central |
手法 | EM (単粒子) / EM (らせん対称) |
解像度 | 3.6 - 5.6 Å |
構造データ | EMDB-4485: EMDB-4487: EMDB-4488: |
由来 |
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