Native phasing of x-ray free-electron laser data for a G protein–coupled receptor

Alexander Batyuk, Lorenzo Galli, Andrii Ishchenko, Gye Won Han, Cornelius Gati, Petr A. Popov, Ming Yue Lee, Benjamin Stauch, Thomas A. White, Anton Barty, Andrew Aquila, Mark S. Hunter, Mengning Liang, Sébastien Boutet, Mengchen Pu, Zhi jie Liu, Garrett Nelson, Daniel James, Chufeng Li, Yun ZhaoJohn C.H. Spence, Wei Liu, Petra Fromme, Vsevolod Katritch, Uwe Weierstall, Raymond C. Stevens, Vadim Cherezov

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52 Citations (Scopus)

Abstract

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 A2A adenosine 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.

Original languageEnglish
Article numbere1600292
JournalScience advances
Volume2
Issue number9
DOIs
Publication statusPublished - Sep 1 2016

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Cite this

Batyuk, A., Galli, L., Ishchenko, A., Han, G. W., Gati, C., Popov, P. A., Lee, M. Y., Stauch, B., White, T. A., Barty, A., Aquila, A., Hunter, M. S., Liang, M., Boutet, S., Pu, M., Liu, Z. J., Nelson, G., James, D., Li, C., ... Cherezov, V. (2016). Native phasing of x-ray free-electron laser data for a G protein–coupled receptor. Science advances, 2(9), [e1600292]. https://doi.org/10.1126/sciadv.1600292