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Proc Natl Acad Sci U S A. 2014 Dec 2;111(48):17122-7. doi: 10.1073/pnas.1418733111. Epub 2014 Oct 31.

Goniometer-based femtosecond crystallography with X-ray free electron lasers.

Author information

1
Stanford Synchrotron Radiation Lightsource, HodgsonK@stanford.edu acohen@slac.stanford.edu.
2
Stanford Synchrotron Radiation Lightsource.
3
Linac Coherent Light Source, and.
4
Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261;
5
Stanford Synchrotron Radiation Lightsource, Departments of Chemistry.
6
Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720;
7
Molecular and Cellular Physiology, and Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305;
8
Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720; Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94158; and.
9
Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59715.
10
Molecular and Cellular Physiology, and.
11
Photon Science, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA 94025; Structural Biology, and.
12
Molecular and Cellular Physiology, and Structural Biology, and.
13
Stanford Synchrotron Radiation Lightsource, Departments of Chemistry, HodgsonK@stanford.edu acohen@slac.stanford.edu.

Abstract

The emerging method of femtosecond crystallography (FX) may extend the diffraction resolution accessible from small radiation-sensitive crystals and provides a means to determine catalytically accurate structures of acutely radiation-sensitive metalloenzymes. Automated goniometer-based instrumentation developed for use at the Linac Coherent Light Source enabled efficient and flexible FX experiments to be performed on a variety of sample types. In the case of rod-shaped Cpl hydrogenase crystals, only five crystals and about 30 min of beam time were used to obtain the 125 still diffraction patterns used to produce a 1.6-Å resolution electron density map. For smaller crystals, high-density grids were used to increase sample throughput; 930 myoglobin crystals mounted at random orientation inside 32 grids were exposed, demonstrating the utility of this approach. Screening results from cryocooled crystals of β2-adrenoreceptor and an RNA polymerase II complex indicate the potential to extend the diffraction resolution obtainable from very radiation-sensitive samples beyond that possible with undulator-based synchrotron sources.

KEYWORDS:

XFEL; crystallography; femtosecond diffraction; structural biology

Comment in

PMID:
25362050
PMCID:
PMC4260607
DOI:
10.1073/pnas.1418733111
[Indexed for MEDLINE]
Free PMC Article

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