Abstract
Serial X-ray crystallography allows macromolecular structure determination at both X-ray free electron lasers (XFELs) and, more recently, synchrotron sources. The time resolution for serial synchrotron crystallography experiments has been limited to millisecond timescales with monochromatic beams. The polychromatic, “pink”, beam provides a more than two orders of magnitude increased photon flux and hence allows accessing much shorter timescales in diffraction experiments at synchrotron sources. Here we report the structure determination of two different protein samples by merging pink-beam diffraction patterns from many crystals, each collected with a single 100 ps X-ray pulse exposure per crystal using a setup optimized for very low scattering background. In contrast to experiments with monochromatic radiation, data from only 50 crystals were required to obtain complete datasets. The high quality of the diffraction data highlights the potential of this method for studying irreversible reactions at sub-microsecond timescales using high-brightness X-ray facilities.
Highlights
Serial X-ray crystallography allows macromolecular structure determination at both X-ray free electron lasers (XFELs) and, more recently, synchrotron sources
Recent advances in highly brilliant X-ray sources, fast framerate detectors and novel sample delivery techniques have changed the way crystallographic data can be collected at both X-ray free electron lasers (XFELs) and third generation synchrotron sources[1,2]
Using our experimental setup and the serial data collection and processing strategy described in the “Methods” section, we collected high-quality diffraction patterns using single-pulse exposures of 100 ps duration per crystal with the pink beam of the BioCARS beamline at the a b
Summary
Serial X-ray crystallography allows macromolecular structure determination at both X-ray free electron lasers (XFELs) and, more recently, synchrotron sources. Owing to the underlying “diffractionbefore-destruction” principle, SFX allows applying orders of magnitude higher doses than in conventional macromolecular X-ray crystallography making it possible to obtain highresolution structural information even from sub-micrometer sized crystals with exposure times in SFX in the 20–50 fs range[4,5,6]. The reduced tolerable dose at room temperature can be compensated by measuring a larger number of crystals. To date, such experiments have been performed with synchrotron radiation monochromatized to a relatively narrow bandwidth of ΔE/E ≈ 10−4. Likewise, increasing the bandwidth reduces the number of snapshots needed for SX, in turn reducing experiment time and sample consumption[11]
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