Serial crystallography on in vivo grown microcrystals using synchrotron radiation

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

doi:10.1107/S2052252513033939

Serial crystallography on in vivo grown microcrystals using synchrotron radiation

IUCrJ. 2014 Feb 10;1(Pt 2):87-94. doi: 10.1107/S2052252513033939. eCollection 2014 Mar 1.

Affiliations

¹ Center for Free-Electron Laser Science (CFEL), Deutsches Elektronensynchrotron (DESY) , Notkestrasse 85, 22607 Hamburg, Germany.
² European Molecular Biology Laboratory (EMBL), Hamburg Outstation , Notkestrasse 85, 22607 Hamburg, Germany.
³ Joint Laboratory for Structural Biology of Infection and Inflammation, Institute of Biochemistry and Molecular Biology, University of Hamburg, and Institute of Biochemistry, University of Lübeck , Notkestrasse 85, 22607 Hamburg, Germany.
⁴ Center for Free-Electron Laser Science (CFEL), Deutsches Elektronensynchrotron (DESY) , Notkestrasse 85, 22607 Hamburg, Germany ; Institute of Biochemistry and Molecular Biology, University of Hamburg , Notkestrasse 85, 22607 Hamburg, Germany.
⁵ Institute of Biochemistry and Molecular Biology, University of Hamburg , Notkestrasse 85, 22607 Hamburg, Germany ; Interfaculty Institute of Biochemistry, University of Tübingen , Hoppe-Seyler-Strasse 4, 72076 Tübingen, Germany.
⁶ Interfaculty Institute of Biochemistry, University of Tübingen , Hoppe-Seyler-Strasse 4, 72076 Tübingen, Germany.
⁷ Institute of Biochemistry and Molecular Biology, University of Hamburg , Notkestrasse 85, 22607 Hamburg, Germany.
⁸ Center for Free-Electron Laser Science (CFEL), Deutsches Elektronensynchrotron (DESY) , Notkestrasse 85, 22607 Hamburg, Germany ; Institute of Experimental Physics, University of Hamburg , Luruper Chaussee 149, 22761 Hamburg, Germany.

Abstract

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 µm(3), 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.

Keywords: in vivo grown microcrystals; protein microcrystallography; serial crystallography.