دورية أكاديمية
In situ serial crystallography facilitates 96-well plate structural analysis at low symmetry
| العنوان: | In situ serial crystallography facilitates 96-well plate structural analysis at low symmetry |
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| المؤلفون: | Nicolas Foos, Jean-Baptise Florial, Mathias Eymery, Jeremy Sinoir, Franck Felisaz, Marcus Oscarsson, Antonia Beteva, Matthew W. Bowler, Didier Nurizzo, Gergely Papp, Montserrat Soler-Lopez, Max Nanao, Shibom Basu, Andrew A. McCarthy |
| المصدر: | IUCrJ, Vol 11, Iss 5, Pp 780-791 (2024) |
| بيانات النشر: | International Union of Crystallography, 2024. |
| سنة النشر: | 2024 |
| المجموعة: | LCC:Crystallography |
| مصطلحات موضوعية: | in situ serial crystallography, room-temperature structures, crystallization plates, conformational flexibility, plate holders, triclinic symmetry, macromolecular crystallography, autotaxin, Crystallography, QD901-999 |
| الوصف: | The advent of serial crystallography has rejuvenated and popularized room-temperature X-ray crystal structure determination. Structures determined at physiological temperature reveal protein flexibility and dynamics. In addition, challenging samples (e.g. large complexes, membrane proteins and viruses) form fragile crystals that are often difficult to harvest for cryo-crystallography. Moreover, a typical serial crystallography experiment requires a large number of microcrystals, mainly achievable through batch crystallization. Many medically relevant samples are expressed in mammalian cell lines, producing a meager quantity of protein that is incompatible with batch crystallization. This can limit the scope of serial crystallography approaches. Direct in situ data collection from a 96-well crystallization plate enables not only the identification of the best diffracting crystallization condition but also the possibility for structure determination under ambient conditions. Here, we describe an in situ serial crystallography (iSX) approach, facilitating direct measurement from crystallization plates mounted on a rapidly exchangeable universal plate holder deployed at a microfocus beamline, ID23-2, at the European Synchrotron Radiation Facility. We applied our iSX approach on a challenging project, autotaxin, a therapeutic target expressed in a stable human cell line, to determine the structure in the lowest-symmetry P1 space group at 3.0 Å resolution. Our in situ data collection strategy provided a complete dataset for structure determination while screening various crystallization conditions. Our data analysis reveals that the iSX approach is highly efficient at a microfocus beamline, improving throughput and demonstrating how crystallization plates can be routinely used as an alternative method of presenting samples for serial crystallography experiments at synchrotrons. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2052-2525 20522525 |
| Relation: | https://journals.iucr.org/paper?S2052252524005785; https://doaj.org/toc/2052-2525 |
| DOI: | 10.1107/S2052252524005785 |
| URL الوصول: | https://doaj.org/article/cf4a521d492f440fa7ef91fbdf26672a |
| رقم الأكسشن: | edsdoj.f4a521d492f440fa7ef91fbdf26672a |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 20522525 |
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| DOI: | 10.1107/S2052252524005785 |