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Dose-resolved serial synchrotron and XFEL structures of radiation-sensitive metalloproteins

DOI: 10.1107/S2052252519003956 DOI Help

Authors: Ali Ebrahim (University of Essex; Diamond Light Source) , Tadeo Moreno-chicano (University of Essex) , Martin V. Appleby (Diamond Light Source) , Amanda K. Chaplin (University of Essex) , John Beale (Diamond Light Source) , Darren A. Sherrell (Argonne National Laboratory) , Helen M. E. Duyvesteyn (Diamond Light Source; University of Oxford) , Shigeki Owada (RIKEN SPring-8 Center; Japan Synchrotron Radiation Research Institute) , Kensuke Tono (RIKEN SPring-8 Center; Japan Synchrotron Radiation Research Institute) , Hiroshi Sugimoto (Japan Synchrotron Radiation Research Institute) , Richard W. Strange (University of Essex) , Jonathan Worrall (University of Essex) , Danny Axford (Diamond Light Source) , Robin L. Owen (Diamond Light Source) , Michael A. Hough (University of Essex)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Iucrj , VOL 6

State: Published (Approved)
Published: July 2019
Diamond Proposal Number(s): 14493

Open Access Open Access

Abstract: An approach is demonstrated to obtain, in a sample- and time-efficient manner, multiple dose-resolved crystal structures from room-temperature protein microcrystals using identical fixed-target supports at both synchrotrons and X-ray free-electron lasers (XFELs). This approach allows direct comparison of dose-resolved serial synchrotron and damage-free XFEL serial femtosecond crystallography structures of radiation-sensitive proteins. Specifically, serial synchrotron structures of a heme peroxidase enzyme reveal that X-ray induced changes occur at far lower doses than those at which diffraction quality is compromised (the Garman limit), consistent with previous studies on the reduction of heme proteins by low X-ray doses. In these structures, a functionally relevant bond length is shown to vary rapidly as a function of absorbed dose, with all room-temperature synchrotron structures exhibiting linear deformation of the active site compared with the XFEL structure. It is demonstrated that extrapolation of dose-dependent synchrotron structures to zero dose can closely approximate the damage-free XFEL structure. This approach is widely applicable to any protein where the crystal structure is altered by the synchrotron X-ray beam and provides a solution to the urgent requirement to determine intact structures of such proteins in a high-throughput and accessible manner.

Journal Keywords: XFELs; microcrystals; serial femtosecond crystallography; serial synchrotron crystallography; serial millisecond crystallography; fixed targets; heme peroxidase; metalloproteins; radiation damage

Subject Areas: Technique Development, Biology and Bio-materials

Diamond Offline Facilities: XFEL Hub
Instruments: I24-Microfocus Macromolecular Crystallography

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