Serial femtosecond crystallography (SFX) takes advantage of extremely bright and ultrashort pulses produced by x-ray free-electron lasers (XFELs), allowing for the collection of high-resolution diffraction intensities from micrometer-sized crystals at room temperature with minimal radiation damage, using the principle of “diffraction-before-destruction.” However, de novo structure factor phase determination using XFELs has been difficult so far. We demonstrate the ability to solve the crystallographic phase problem for SFX data collected with an XFEL using the anomalous signal from native sulfur atoms, leading to a bias-free room temperature structure of the human A2Aadenosine receptor at 1.9 Å resolution. The advancement was made possible by recent improvements in SFX data analysis and the design of injectors and delivery media for streaming hydrated microcrystals. This general method should accelerate structural studies of novel difficult-to-crystallize macromolecules and their complexes.
Batyuk, Alexander, et al. "Native phasing of x-ray free-electron laser data for a G protein–coupled receptor." Science Advances, vol. 2, no. 9, Sep. 2016. https://doi.org/10.1126/sciadv.1600292
Batyuk, Alexander, Galli, Lorenzo, Ishchenko, Andrii, et al., "Native phasing of x-ray free-electron laser data for a G protein–coupled receptor," Science Advances 2, no. 9 (2016), https://doi.org/10.1126/sciadv.1600292
@article{osti_1625958,
author = {Batyuk, Alexander and Galli, Lorenzo and Ishchenko, Andrii and Han, Gye Won and Gati, Cornelius and Popov, Petr A. and Lee, Ming-Yue and Stauch, Benjamin and White, Thomas A. and Barty, Anton and others},
title = {Native phasing of x-ray free-electron laser data for a G protein–coupled receptor},
annote = {Serial femtosecond crystallography (SFX) takes advantage of extremely bright and ultrashort pulses produced by x-ray free-electron lasers (XFELs), allowing for the collection of high-resolution diffraction intensities from micrometer-sized crystals at room temperature with minimal radiation damage, using the principle of “diffraction-before-destruction.” However, de novo structure factor phase determination using XFELs has been difficult so far. We demonstrate the ability to solve the crystallographic phase problem for SFX data collected with an XFEL using the anomalous signal from native sulfur atoms, leading to a bias-free room temperature structure of the human A2Aadenosine receptor at 1.9 Å resolution. The advancement was made possible by recent improvements in SFX data analysis and the design of injectors and delivery media for streaming hydrated microcrystals. This general method should accelerate structural studies of novel difficult-to-crystallize macromolecules and their complexes.},
doi = {10.1126/sciadv.1600292},
url = {https://www.osti.gov/biblio/1625958},
journal = {Science Advances},
issn = {ISSN 2375-2548},
number = {9},
volume = {2},
place = {United States},
publisher = {AAAS},
year = {2016},
month = {09}}
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH); Ministry of Science and Technology of China; National Science Foundation (NSF), Science and Technology Center; PIER Helmholtz Graduate School; Helmholtz Association; Russian Ministry of Education and Science
Coherent X-ray Imaging Data Bank (Lawrence Berkeley National Laboratory); The University of Tokyo, RIKEN SPring-8 Center, POSTECH, Osaka University, Kyoto University, Japan Synchrotron Radiation Research Institutehttps://doi.org/10.11577/1274312