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High-throughput structures of protein–ligand complexes at room temperature using serial femtosecond crystallography

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). 1074 - 1085. ISSN 2052-2525

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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
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 > Life Sciences, School of
Depositing User: Elements
Date Deposited: 22 Nov 2019 10:44
Last Modified: 22 Nov 2019 11:15
URI: http://repository.essex.ac.uk/id/eprint/25988

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