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

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)...

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Detalles Bibliográficos
Autores principales: Moreno-Chicano, Tadeo, Ebrahim, Ali, Axford, Danny, Appleby, Martin V., Beale, John H., Chaplin, Amanda K., Duyvesteyn, Helen M. E., Ghiladi, Reza A., Owada, Shigeki, Sherrell, Darren A., Strange, Richard W., Sugimoto, Hiroshi, Tono, Kensuke, Worrall, Jonathan A. R., Owen, Robin L., Hough, Michael A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830213/
https://www.ncbi.nlm.nih.gov/pubmed/31709063
http://dx.doi.org/10.1107/S2052252519011655
Descripción
Sumario: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.