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

Ebrahim, Ali and Moreno-Chicano, Tadeo and Appleby, Martin V and Chaplin, Amanda K and Beale, John H and Sherrell, Darren A and Duyvesteyn, Helen ME and Owada, Shigeki and Tono, Kensuke and Sugimoto, Hiroshi and Strange, Richard W and Worrall, Jonathan AR and Axford, Danny and Owen, Robin L and Hough, Michael A (2019) 'Dose-resolved serial synchrotron and XFEL structures of radiation-sensitive metalloproteins.' IUCrJ, 6 (4). 543 - 551. ISSN 2052-2525

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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.

Item Type: Article
Divisions: Faculty of Science and Health > Life Sciences, School of
Depositing User: Elements
Date Deposited: 25 Sep 2019 15:40
Last Modified: 25 Sep 2019 16:15
URI: http://repository.essex.ac.uk/id/eprint/25309

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