Moreno-Chicano, Tadeo and Ebrahim, Ali and Axford, Danny and Appleby, Martin V and Beale, John H and Chaplin, Amanda K and Duyvesteyn, Helen ME and Ghiladi, Reza A and Owada, Shigeki and Sherrell, Darren A and Strange, Richard W and Sugimoto, Hiroshi and Tono, Kensuke and Worrall, Jonathan AR and Owen, Robin L and Hough, Michael A (2019) High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography. IUCrJ, 6 (6). pp. 1074-1085. DOI https://doi.org/10.1107/s2052252519011655
Moreno-Chicano, Tadeo and Ebrahim, Ali and Axford, Danny and Appleby, Martin V and Beale, John H and Chaplin, Amanda K and Duyvesteyn, Helen ME and Ghiladi, Reza A and Owada, Shigeki and Sherrell, Darren A and Strange, Richard W and Sugimoto, Hiroshi and Tono, Kensuke and Worrall, Jonathan AR and Owen, Robin L and Hough, Michael A (2019) High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography. IUCrJ, 6 (6). pp. 1074-1085. DOI https://doi.org/10.1107/s2052252519011655
Moreno-Chicano, Tadeo and Ebrahim, Ali and Axford, Danny and Appleby, Martin V and Beale, John H and Chaplin, Amanda K and Duyvesteyn, Helen ME and Ghiladi, Reza A and Owada, Shigeki and Sherrell, Darren A and Strange, Richard W and Sugimoto, Hiroshi and Tono, Kensuke and Worrall, Jonathan AR and Owen, Robin L and Hough, Michael A (2019) High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography. IUCrJ, 6 (6). pp. 1074-1085. DOI https://doi.org/10.1107/s2052252519011655
Abstract
High-throughput X-ray crystal structures of protein–ligand complexes are critical to pharmaceutical drug development. However, cryocooling of crystals and X-ray radiation damage may distort the observed ligand binding. Serial femtosecond crystallography (SFX) using X-ray free-electron lasers (XFELs) can produce radiation-damage-free room-temperature structures. Ligand-binding studies using SFX have received only modest attention, partly owing to limited beamtime availability and the large quantity of sample that is required per structure determination. Here, a high-throughput approach to determine room-temperature damage-free structures with excellent sample and time efficiency is demonstrated, allowing complexes to be characterized rapidly and without prohibitive sample requirements. This yields high-quality difference density maps allowing unambiguous ligand placement. Crucially, it is demonstrated that ligands similar in size or smaller than those used in fragment-based drug design may be clearly identified in data sets obtained from <1000 diffraction images. This efficiency in both sample and XFEL beamtime opens the door to true high-throughput screening of protein–ligand complexes using SFX.
Item Type: | Article |
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Uncontrolled Keywords: | serial femtosecond crystallography; ligand binding; high throughput; X-ray crystallography; damage-free structures; X-ray free-electron lasers |
Divisions: | Faculty of Science and Health Faculty of Science and Health > Life Sciences, School of |
SWORD Depositor: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 22 Nov 2019 10:44 |
Last Modified: | 07 Aug 2024 20:43 |
URI: | http://repository.essex.ac.uk/id/eprint/25988 |
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Filename: MorenoChicano2019.pdf
Licence: Creative Commons: Attribution 3.0