Selenium single-wavelength anomalous diffraction de novo phasing using an X-ray-free electron laser.
Structural information about biological macromolecules near the atomic scale provides important insight into the functions of these molecules. To date, X-ray crystallography has been the predominant method used for macromolecular structure determination. However, challenges exist when solving structures with X-rays, including the phase problem and radiation damage. X-ray-free electron lasers (X-ray FELs) have enabled collection of diffraction information before the onset of radiation damage, yet the majority of structures solved at X-ray FELs have been phased using external information via molecular replacement. De novo phasing at X-ray FELs has proven challenging due in part to per-pulse variations in intensity and wavelength. Here we report the solution of a selenobiotinyl-streptavidin structure using phases obtained by the anomalous diffraction of selenium measured at a single wavelength (Se-SAD) at the Linac Coherent Light Source. Our results demonstrate Se-SAD, routinely employed at synchrotrons for novel structure determination, is now possible at X-ray FELs.
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Mark S
MS
Hunter
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Chun Hong
CH
Yoon
Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Hasan
0000-0002-9135-5397
H
DeMirci
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Raymond G
RG
Sierra
Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
E Han
EH
Dao
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Radman
R
Ahmadi
Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Fulya
F
Aksit
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Andrew L
AL
Aquila
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Halilibrahim
H
Ciftci
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Serge
S
Guillet
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Matt J
MJ
Hayes
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Thomas J
TJ
Lane
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Meng
M
Liang
Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Ulf
U
Lundström
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Jason E
0000-0001-6811-6083
JE
Koglin
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Paul
P
Mgbam
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Yashas
Y
Rao
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Lindsey
L
Zhang
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Soichi
S
Wakatsuki
Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.|Department of Biochemistry and Biophysics, University of California, San Francisco, California 94158, USA.
James M
JM
Holton
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
Sébastien
S
Boutet
D001426:Bacterial Proteins / Q000737:chemistry / Q000648:ultrastructure*; D001710:Biotin / Q000031:analogs & derivatives* / Q000737:chemistry; D018360:Crystallography, X-Ray / Q000295:instrumentation / Q000379:methods*; D004583:Electrons; D005240:Feasibility Studies; D007834:Lasers; D008958:Models, Molecular; D011487:Protein Conformation; D012643:Selenium / Q000737:chemistry*
D001426:Bacterial Proteins; C065877:selenobiotinyl streptavidin; D001710:Biotin; D012643:Selenium