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

DOI: 10.1107/S2052252519011655 DOI Help

Authors: Tadeo Moreno Chicano (University of Essex) , Ali Ebrahim (University of Essex) , Danny Axford (Diamond Light Source) , Martin V. Appleby (Diamond Light Source) , John H. Beale (Diamond Light Source) , Amanda K. Chaplin (University of Essex) , Helen M. E. Duyvesteyn (University of Oxford; Diamond Light Source) , Reza A. Ghiladi (North Carolina State University) , Shigeki Owada (RIKEN SPring-8 Center; Japan Synchrotron Radiation Research Institute) , Darren A. Sherrell (Diamond Light Source) , Richard Strange (University of Essex) , Hiroshi Sugimoto (RIKEN SPring-8 Center) , Kensuke Tono (RIKEN SPring-8 Center; Japan Synchrotron Radiation Research Institute) , Jonathan A. R. Worrall (University of Essex) , 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: November 2019

Open Access Open Access

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.

Journal Keywords: serial femtosecond crystallography; ligand binding; high throughput; X-ray crystallography; damage-free structures; X-ray free-electron lasers

Subject Areas: Biology and Bio-materials, Medicine, Technique Development


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