| Autor: |
Voss, Jonathan M., Harder, Oliver F., Olshin, Pavel K., Drabbels, Marcel, Lorenz, Ulrich J. |
| Rok vydání: |
2021 |
| Předmět: |
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| Druh dokumentu: |
Working Paper |
| DOI: |
10.1016/j.cplett.2021.138812 |
| Popis: |
Proteins typically undergo conformational dynamics on the microsecond to millisecond timescale as they perform their function, which is much faster than the time-resolution of cryo-electron microscopy and has thus prevented real-time observations. Here, we propose a novel approach for microsecond time-resolved cryo-electron microscopy that involves rapidly melting a cryo specimen in situ with a laser beam. The sample remains liquid for the duration of the laser pulse, offering a tunable time window in which the dynamics of embedded particles can be induced in their native liquid environment. After the laser pulse, the sample vitrifies in just a few microseconds, trapping particles in their transient configurations, so that they can subsequently be characterized with conventional cryo-electron microscopy. We demonstrate that our melting and revitrification approach is viable and affords microsecond time resolution. As a proof of principle, we study the disassembly of particles after they incur structural damage and trap them in partially unraveled configurations. |
| Databáze: |
arXiv |
| Externí odkaz: |
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