Serial crystallography on in vivo grown microcrystals using synchrotron radiation

• Main Authors: Cornelius Gati, Gleb Bourenkov, Marco Klinge, Dirk Rehders, Francesco Stellato, Dominik Oberthür, Oleksandr Yefanov, Benjamin P. Sommer, Stefan Mogk, Michael Duszenko, Christian Betzel, Thomas R. Schneider, Henry N. Chapman, Lars Redecke
• Format: Article
• Language: English
• Published: International Union of Crystallography 2014-03-01
• Series: IUCrJ
• Subjects: protein microcrystallography; serial crystallography; in vivo grown microcrystals
• Online Access: http://scripts.iucr.org/cgi-bin/paper?S2052252513033939

Crystal structure determinations of biological macromolecules are limited by the availability of sufficiently sized crystals and by the fact that crystal quality deteriorates during data collection owing to radiation damage. Exploiting a micrometre-sized X-ray beam, high-precision diffractometry and shutterless data acquisition with a pixel-array detector, a strategy for collecting data from many micrometre-sized crystals presented to an X-ray beam in a vitrified suspension is demonstrated. By combining diffraction data from 80 Trypanosoma brucei procathepsin B crystals with an average volume of 9 µm3, a complete data set to 3.0 Å resolution has been assembled. The data allowed the refinement of a structural model that is consistent with that previously obtained using free-electron laser radiation, providing mutual validation. Further improvements of the serial synchrotron crystallography technique and its combination with serial femtosecond crystallography are discussed that may allow the determination of high-resolution structures of micrometre-sized crystals.