High-Resolution Protein Structure Determination by Serial Femtosecond Crystallography

By S. Boutet, L. Lomb, Garth J. Williams, T. Barends, A. Aquila, R. Doak, U. Weierstall, D. DePonte, J. Steinbrener, R. Shoeman and 44 more. Science, 2012

820 citations recorded by the source corpus.

Size Matters Less X-ray crystallography is a central research tool for uncovering the structures of proteins and other macromolecules. However, its applicability typically requires growth of large crystals, in part because a sufficient number of molecules must be present in the lattice for the sample to withstand x-ray—induced damage. Boutet et al. (p. 362, published online 31 May) now demonstrate that the intense x-ray pulses emitted by a free-electron laser source can yield data in few enough exposures to uncover the high-resolution structure of microcrystals. A powerful x-ray laser source can probe proteins in detail using much smaller crystals than previously required. Structure determination of proteins and other macromolecules has historically required the growth of high-quality crystals sufficiently large to diffract x-rays efficiently while withstanding radiation damage. We applied serial femtosecond crystallography (SFX) using an x-ray free-electron laser (XFEL) to obtain high-resolution structural information from microcrystals (less than 1 micrometer by 1 micrometer by 3 micrometers) of the well-characterized model protein lysozyme. The agreement with synchrotron data demonstrates the immediate relevance of SFX for analyzing the structure of the large group of difficult-to-crystallize molecules.

Subfields: Materials Chemistry, Radiation, Structural Biology

View Paper: https://www.semanticscholar.org/paper/fbe26feab9768303bf79c9ef4559a3b92ab17bb8

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Citations in the corpus

A glimpse of structural biology through X-ray crystallography. (2014, background): “The first high-resolution X-ray structure by serial femtosecond crystallography (SFX) was accomplished on lysozyme (Boutet et al., 2012), followed by de novo generation of experimental phases for lysozyme (Barends et al., 2014).”
10 years of pioneering X-ray science at the Free-Electron Laser FLASH at DESY (2019, background): “Although the first high-resolution (2 Å) structures were only demonstrated a few years ago (Boutet et al., 2012), this technique is already having significant impact on structural biology through the study of hard-to-crystallize but medically relevant G protein-coupled receptors (GPCR) at room…”
Cheetah: software for high-throughput reduction and analysis of serial femtosecond X-ray diffraction data (2014, background): “Data presented in this paper formed part of the data presented by Boutet et al. (2012).”
Serial Synchrotron X-Ray Crystallography (SSX). (2017, neutral): “2–2 μm) [13], lysozyme (1 × 1 × 3 μm3) [20], and cathepsin B (0.”
Serum Albumin Domain Structures in Human Blood Serum by Mass Spectrometry and Computational Biology (2015, background): “New developments to overcome some of these restrictions include x-ray free electron lasers (XFEL) (4), which only require microcrystals, new detectors in cryo-electron microscopy (5) and in-cell NMR (6), which analyzes the structure of small proteins in a cellular context.”
SwissFEL: The Swiss X-ray Free Electron Laser (2017, methodology): “SFX [19] is an XFEL technique that has been developed for protein crystallography where a stream of micrometer-sized crystals is delivered into the focussed X-ray beam using a range of different injector techniques [135–137], and the diffraction pattern from each crystal is recorded on a large two-d”
Serial femtosecond crystallography: the first five years (2015, background): “A comparison of highresolution SFX and conventional rotation data sets collected at the LCLS and Swiss Light Source, respectively, showed good agreement of both the diffraction intensities and their statistics, as well as of the refined models (Boutet et al., 2012).”