Native phasing of x-ray free-electron laser data for a G protein-coupled receptor.
Alexander BatyukLorenzo GalliAndrii IshchenkoGye Won HanCornelius GatiPetr A PopovMing-Yue LeeBenjamin StauchThomas A WhiteAnton BartyAndrew AquilaMark S HunterMengning LiangSébastien BoutetMengchen PuZhi-Jie LiuGarrett NelsonDaniel JamesChufeng LiYun ZhaoJohn C H SpenceWei LiuPetra FrommeVsevolod KatritchUwe WeierstallRaymond C StevensVadim CherezovPublished in: Science advances (2016)
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.
Keyphrases
- room temperature
- data analysis
- high resolution
- electron microscopy
- ionic liquid
- electronic health record
- endothelial cells
- big data
- oxidative stress
- dual energy
- computed tomography
- high speed
- crystal structure
- magnetic resonance
- radiation induced
- induced pluripotent stem cells
- binding protein
- artificial intelligence
- pluripotent stem cells
- simultaneous determination
- protein kinase