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 Zhao; John Spence; Wei Liu; Petra Fromme; Vsevolod Katritch; Uwe Weierstall; Raymond C. Stevens; Vadim Cherezov

doi:10.1126/sciadv.1600292

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 Spence, Wei Liu, Petra Fromme, Vsevolod Katritch, Uwe Weierstall, Raymond C. Stevens, Vadim Cherezov

Research output: Contribution to journal › Article › peer-review

96 Scopus citations

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 A_2A 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 language	English (US)
Article number	e1600292
Journal	Science Advances
Volume	2
Issue number	9
DOIs	https://doi.org/10.1126/sciadv.1600292
State	Published - Sep 2016

ASJC Scopus subject areas

General

Access to Document

10.1126/sciadv.1600292

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), Article e1600292. https://doi.org/10.1126/sciadv.1600292

Batyuk, A, Galli, L, Ishchenko, A, Han, GW, Gati, C, Popov, PA, Lee, MY, Stauch, B, White, TA, Barty, A, Aquila, A, Hunter, MS, Liang, M, Boutet, S, Pu, M, Liu, ZJ, Nelson, G, James, D, Li, C, Zhao, Y, Spence, J, Liu, W, Fromme, P, Katritch, V, Weierstall, U, Stevens, RC & Cherezov, V 2016, 'Native phasing of x-ray free-electron laser data for a G protein–coupled receptor', Science Advances, vol. 2, no. 9, e1600292. https://doi.org/10.1126/sciadv.1600292

@article{a911d47ddccf4c718d4a5ae89a9dc0e8,

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

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 {\AA} 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.",

author = "Alexander Batyuk and Lorenzo Galli and Andrii Ishchenko and Han, {Gye Won} and Cornelius Gati and Popov, {Petr A.} and Lee, {Ming Yue} and Benjamin Stauch and White, {Thomas A.} and Anton Barty and Andrew Aquila and Hunter, {Mark S.} and Mengning Liang and S{\'e}bastien Boutet and Mengchen Pu and Liu, {Zhi jie} and Garrett Nelson and Daniel James and Chufeng Li and Yun Zhao and John Spence and Wei Liu and Petra Fromme and Vsevolod Katritch and Uwe Weierstall and Stevens, {Raymond C.} and Vadim Cherezov",

note = "Publisher Copyright: 2016 {\textcopyright} The Authors.",

year = "2016",

month = sep,

doi = "10.1126/sciadv.1600292",

language = "English (US)",

volume = "2",

journal = "Science Advances",

issn = "2375-2548",

publisher = "American Association for the Advancement of Science",

number = "9",

}

TY - JOUR

T1 - Native phasing of x-ray free-electron laser data for a G protein–coupled receptor

AU - Batyuk, Alexander

AU - Galli, Lorenzo

AU - Ishchenko, Andrii

AU - Han, Gye Won

AU - Gati, Cornelius

AU - Popov, Petr A.

AU - Lee, Ming Yue

AU - Stauch, Benjamin

AU - White, Thomas A.

AU - Barty, Anton

AU - Aquila, Andrew

AU - Hunter, Mark S.

AU - Liang, Mengning

AU - Boutet, Sébastien

AU - Pu, Mengchen

AU - Liu, Zhi jie

AU - Nelson, Garrett

AU - James, Daniel

AU - Li, Chufeng

AU - Zhao, Yun

AU - Spence, John

AU - Liu, Wei

AU - Fromme, Petra

AU - Katritch, Vsevolod

AU - Weierstall, Uwe

AU - Stevens, Raymond C.

AU - Cherezov, Vadim

PY - 2016/9

Y1 - 2016/9

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85042167986&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042167986&partnerID=8YFLogxK

U2 - 10.1126/sciadv.1600292

DO - 10.1126/sciadv.1600292

M3 - Article

AN - SCOPUS:85042167986

SN - 2375-2548

VL - 2

JO - Science Advances

JF - Science Advances

IS - 9

M1 - e1600292

ER -

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

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

1.9A angstrom A2a adenosine receptor structure with MR phasing using XFEL data

2.5 angstrom A2a adenosine receptor structure with MR phasing using XFEL data

2.5 angstrom A2a adenosine receptor structure with sulfur SAD phasing using XFEL data

Cite this

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

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Datasets

1.9A angstrom A2a adenosine receptor structure with MR phasing using XFEL data

2.5 angstrom A2a adenosine receptor structure with MR phasing using XFEL data

2.5 angstrom A2a adenosine receptor structure with sulfur SAD phasing using XFEL data

Cite this