Time-resolved macromolecular crystallography (TR-MX) at synchrotrons has long been limited to the use of the Laue diffraction technique and was only successful for a handful of biological systems. The recent advent of X-ray free-electron lasers has rejuvenated the field by fostering the development of serial crystallography, which consists in composing a complete data set from single diffraction images obtained from tens of thousands of microcrystals passing through a pulsed X-ray beam. The associated development of crystal injection techniques, coupled to the advent of faster, noiseless X-ray detectors, has paved the way for the revival of TR-MX at synchrotrons. The EBSL8/ID29 beamline (beamline responsible, Daniele de Sanctis) currently under construction as part of the ESRF EBS Upgrade Project, has been designed to offer a variety of techniques and methods for TR-MX with a microsecond time resolution. Additionally, the ESRF Structural Biology Group has developed an oscillation-based TR-MX method, demonstrating its applicability with the study of a photoreceptor fragment at a time resolution of 63 ms (Aumonier et al., IUCrJ (2020)). We propose to study this target, as well as other photoreceptor fragments and photosensitive proteins, by various TR-MX methods (fixed-target-; grease injector-; tape-drive- or oscillation-based) to identify those which are most suitable for further development on EBSL8/ID29 and to make molecular movies at the lowest time resolution attainable, ideally 100 μs. This work will be performed in co-supervision with Prof. Arwen Pearson (CFEL, University of Hamburg, Germany) with whom previously developed methods for TR-MX will be evaluated using beamlines both at the ESRF and PETRA III, most particularly at the T-REXX endstation on beamline P14, operated by EMBL-Hamburg (Mehrabi et al., Nat. Methods (2019)).
Further information may be obtained from Antoine ROYANT (tel.: +33 (0)4 76 88 17 46, email: firstname.lastname@example.org).
Contract of two years employed by the ESRF, with a monthly gross salary of 2339 €, then of two years employed by the University of Hamburg.
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