Abstract
Owing to the development of brilliant microfocus beamlines, rapid-readout detectors and sample changers, protein microcrystallography is rapidly becoming a popular technique for accessing structural information from complex biological samples. However, the method is time-consuming and labor-intensive and requires technical expertise to obtain high-resolution protein crystal structures. At SPring-8, an automated data-collection system named ZOO has been developed. This system enables faster data collection, facilitates advanced data-collection and data-processing techniques, and permits the collection of higher quality data. In this paper, the key features of the functionality put in place on the SPring-8 microbeam beamline BL32XU are described and the major advantages of this system are outlined. The ZOO system will be a major driving force in the evolution of the macromolecular crystallography beamlines at SPring-8.
Highlights
The elucidation of high-resolution structures of biological macromolecules has contributed greatly to our understanding of biological processes at the molecular level
For crystals grown by the in meso method, the multiple small-wedge scheme, which was originally reported for the high-resolution structural analysis of G-proteincoupled receptors (GPCRs), is a useful approach that can be performed at cryogenic temperatures (Cherezov et al, 2007; Rasmussen et al, 2012; Rosenbaum et al, 2012)
We describe an automated data-collection system, ZOO, and evaluate its ability to analyze various experimental schemes
Summary
The elucidation of high-resolution structures of biological macromolecules has contributed greatly to our understanding of biological processes at the molecular level. For crystals grown by the in meso method, the multiple small-wedge scheme, which was originally reported for the high-resolution structural analysis of G-proteincoupled receptors (GPCRs), is a useful approach that can be performed at cryogenic temperatures (Cherezov et al, 2007; Rasmussen et al, 2012; Rosenbaum et al, 2012) In this scheme, X-ray-based raster scanning is applied to a cryoloop containing several tens of crystals to specify each crystal position (Cherezov et al, 2009). The ZOO system has automated all conventional goniometerbased data-collection schemes, such as the normal rotation method, helical data collection, multiple small-wedge and synchrotron serial rotation crystallography under cryogenic conditions. We describe how ZOO works and how ZOO can contribute to challenging structure analyses
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