Serial millisecond crystallography for routine room-temperature structure determination at synchrotrons

• Published in: Nature communications 2017-09, Vol.8 (1), p.542-11, Article 542
• Main Authors: Weinert, Tobias, Olieric, Natacha, Cheng, Robert, Brünle, Steffen, James, Daniel, Ozerov, Dmitry, Gashi, Dardan, Vera, Laura, Marsh, May, Jaeger, Kathrin, Dworkowski, Florian, Panepucci, Ezequiel, Basu, Shibom, Skopintsev, Petr, Doré, Andrew S., Geng, Tian, Cooke, Robert M., Liang, Mengning, Prota, Andrea E., Panneels, Valerie, Nogly, Przemyslaw, Ermler, Ulrich, Schertler, Gebhard, Hennig, Michael, Steinmetz, Michel O., Wang, Meitian, Standfuss, Jörg
• Format: Article
• Language: English
• Subjects: 631/154; 631/45/535/1266; Adenosine; BASIC BIOLOGICAL SCIENCES; Colchicine; Collection; Crystal structure; Crystallography; Data acquisition; Data collection; Diffraction; Diffraction patterns; Drug discovery; Free electron lasers; High resolution; Historical account; Humanities and Social Sciences; Injectors; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Lasers; Molybdenum; multidisciplinary; Proteins; Radiation; Radiation damage; Room temperature; Science; Science (multidisciplinary); Structural damage; Sulfur; Synchrotrons; Temperature effects; Tubulin; Viscosity; X-ray crystallography; X-rays
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-017-00630-4

Historically, room-temperature structure determination was succeeded by cryo-crystallography to mitigate radiation damage. Here, we demonstrate that serial millisecond crystallography at a synchrotron beamline equipped with high-viscosity injector and high frame-rate detector allows typical crystallographic experiments to be performed at room-temperature. Using a crystal scanning approach, we determine the high-resolution structure of the radiation sensitive molybdenum storage protein, demonstrate soaking of the drug colchicine into tubulin and native sulfur phasing of the human G protein-coupled adenosine receptor. Serial crystallographic data for molecular replacement already converges in 1,000–10,000 diffraction patterns, which we collected in 3 to maximally 82 minutes. Compared with serial data we collected at a free-electron laser, the synchrotron data are of slightly lower resolution, however fewer diffraction patterns are needed for de novo phasing. Overall, the data we collected by room-temperature serial crystallography are of comparable quality to cryo-crystallographic data and can be routinely collected at synchrotrons. Serial crystallography was developed for protein crystal data collection with X-ray free-electron lasers. Here the authors present several examples which show that serial crystallography using high-viscosity injectors can also be routinely employed for room-temperature data collection at synchrotrons.