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-Structure paper
タイトル | Microsecond melting and revitrification of cryo samples: protein structure and beam-induced motion. |
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ジャーナル・号・ページ | Acta Crystallogr D Struct Biol, Vol. 78, Issue Pt 7, Page 883-889, Year 2022 |
掲載日 | 2022年7月1日 |
著者 | Oliver F Harder / Jonathan M Voss / Pavel K Olshin / Marcel Drabbels / Ulrich J Lorenz / |
PubMed 要旨 | A novel approach to time-resolved cryo-electron microscopy (cryo-EM) has recently been introduced that involves melting a cryo sample with a laser beam to allow protein dynamics to briefly occur in ...A novel approach to time-resolved cryo-electron microscopy (cryo-EM) has recently been introduced that involves melting a cryo sample with a laser beam to allow protein dynamics to briefly occur in the liquid, before trapping the particles in their transient configurations by rapidly revitrifying the sample. With a time resolution of just a few microseconds, this approach is notably fast enough to study the domain motions that are typically associated with the activity of proteins but which have previously remained inaccessible. Here, crucial details are added to the characterization of the method. It is shown that single-particle reconstructions of apoferritin and Cowpea chlorotic mottle virus from revitrified samples are indistinguishable from those from conventional samples, demonstrating that melting and revitrification leaves the particles intact and that they do not undergo structural changes within the spatial resolution afforded by the instrument. How rapid revitrification affects the properties of the ice is also characterized, showing that revitrified samples exhibit comparable amounts of beam-induced motion. The results pave the way for microsecond time-resolved studies of the conformational dynamics of proteins and open up new avenues to study the vitrification process and to address beam-induced specimen movement. |
リンク | Acta Crystallogr D Struct Biol / PubMed:35775987 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 4.25 - 5.2 Å |
構造データ | EMDB-14767: Mouse heavy chain apoferritin in plunge-frozen vitrified ice EMDB-14768: Mouse heavy chain apoferritin in laser-melted and revitrified ice EMDB-14769: Cowpea chlorotic mottle virus in plunge-frozen ice EMDB-14772: Cowpea chlorotic mottle virus in laser-melted and revitrified ice |
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