Native phasing of x-ray free-electron laser data for a G protein-coupled receptor.
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.
Department of Biochemistry, University of Zürich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
Alexander
0000-0002-9393-2880
A
Batyuk
Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Lorenzo
L
Galli
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
Andrii
A
Ishchenko
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
Gye Won
0000-0003-4107-835X
GW
Han
Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Cornelius
C
Gati
Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.; Moscow Institute of Physics and Technology, Dolgoprudny 141700, Russia.; Department of Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA.
Petr A
PA
Popov
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
Ming-Yue
0000-0001-7082-1216
MY
Lee
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
Benjamin
0000-0001-7626-2021
B
Stauch
Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Thomas A
0000-0002-8407-0270
TA
White
Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
Anton
A
Barty
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Andrew
A
Aquila
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Mark S
0000-0002-0110-7075
MS
Hunter
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Mengning
M
Liang
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Sébastien
S
Boutet
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
Mengchen
0000-0001-6282-2454
M
Pu
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.; iHuman Institute, ShanghaiTech University, Shanghai 201210, China.
Zhi-Jie
0000-0001-7279-2893
ZJ
Liu
Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
Garrett
G
Nelson
Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
Daniel
0000-0002-8348-6661
D
James
Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
Chufeng
0000-0002-2997-0903
C
Li
Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
Yun
0000-0003-4559-399X
Y
Zhao
Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
John C H
JC
Spence
Center for Applied Structural Discovery at the Biodesign Institute, School of Molecular Sciences, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA.
Wei
W
Liu
Center for Applied Structural Discovery at the Biodesign Institute, School of Molecular Sciences, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA.
Petra
P
Fromme
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.; Department of Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA.
Vsevolod
V
Katritch
Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
Uwe
U
Weierstall
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.; Department of Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA.; iHuman Institute, ShanghaiTech University, Shanghai 201210, China.
Raymond C
RC
Stevens
Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.; Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.; Moscow Institute of Physics and Technology, Dolgoprudny 141700, Russia.; Department of Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA.; Department of Physics and Astronomy, University of Southern California, Los Angeles, CA 90089, USA.
Vadim
V
Cherezov
Crystallography; GPCR; SAD; de novo structure; lipidic cubic phase; native phasing; protein; serial femtosecond crystallography; sulfur; x-ray free electron laser
J Struct Biol. 2004 Nov;148(2):169-75
15477097
IUCrJ. 2015 Feb 03;2(Pt 2):246-55
25866661
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
20383002
J Synchrotron Radiat. 2015 May;22(3):514-9
25931062
Sci Rep. 2015 Sep 11;5:14017
26360462
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
19461840
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
20057044
Trends Pharmacol Sci. 2012 May;33(5):249-60
22465153
Acta Crystallogr D Biol Crystallogr. 2012 Apr;68(Pt 4):352-67
22505256
Cell. 2015 May 7;161(4):833-44
25913193
Acta Crystallogr D Biol Crystallogr. 2014 Aug;70(Pt 8):2204-16
25084339
Acta Crystallogr D Biol Crystallogr. 2015 Dec 1;71(Pt 12):2519-25
26627659
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):479-85
20383001
Philos Trans R Soc Lond B Biol Sci. 2014 Jul 17;369(1647):20130330
24914157
Nature. 1981 Mar 12;290(5802):107-113
28769131
Nature. 2015 Jul 30;523(7562):561-7
26200343
Acta Crystallogr A. 2011 Mar;67(Pt 2):131-40
21325716
IUCrJ. 2016 Mar 09;3(Pt 3):180-91
27158504
Elife. 2015 Mar 17;4:null
25781634
Nature. 2014 Jan 9;505(7482):244-7
24270807
Trends Biochem Sci. 2014 May;39(5):233-44
24767681
IUCrJ. 2015 Aug 04;2(Pt 5):545-51
26306196
Acta Crystallogr D Biol Crystallogr. 2003 Jun;59(Pt 6):1020-7
12777765
Nat Methods. 2015 Jan;12(1):61-3
25384243
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
20124702
Science. 2013 Dec 20;342(6165):1521-4
24357322
Nat Protoc. 2009;4(5):706-31
19390528
Nat Methods. 2015 Feb;12(2):131-3
25506719
Science. 2012 Jul 13;337(6091):232-6
22798613
J Synchrotron Radiat. 2015 May;22(3):577-83
25931071
Nat Protoc. 2014 Sep;9(9):2123-34
25122522
IUCrJ. 2015 Jun 30;2(Pt 4):421-30
26177184
Nat Commun. 2014;5:3309
24525480
Proc Natl Acad Sci U S A. 2016 Mar 15;113(11):2928-33
26929369
Acta Crystallogr D Biol Crystallogr. 2005 May;61(Pt 5):520-7
15858261
Acta Crystallogr D Biol Crystallogr. 2000 Aug;56(Pt 8):965-72
10944333
Nature. 2011 Feb 3;470(7332):73-7
21293373
J Appl Crystallogr. 2014 May 29;47(Pt 3):1118-1131
24904246
Science. 2014 Dec 5;346(6214):1242-6
25477465
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32
20124692
Science. 2012 May 25;336(6084):1033-7
22628655
Opt Express. 2015 Nov 2;23 (22):28459-70
26561117