期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 111, 期 48, 页码 17122-17127出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1418733111
关键词
femtosecond diffraction; crystallography; XFEL; structural biology
资金
- US Department of Energy, Office of Basic Energy Sciences
- Use of the Stanford Synchrotron Radiation Lightsource (SSRL)
- SLAC National Accelerator Laboratory
- US Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-76SF00515]
- US Department of Energy Office of Biological and Environmental Research
- National Institutes of Health (NIH)
- National Institute of General Medical Sciences [P41GM103393]
- Howard Hughes Medical Institute Collaborative Innovation Award
- NIH [GM095887, GM102520, GM073210, GM082250, GM094625]
- Biological and Electron Transfer
- Catalysis EFRC
- US Department of Energy, Office of Science [DE-SC0012518]
- U.S. Department of Energy (DOE) [DE-SC0012518] Funding Source: U.S. Department of Energy (DOE)
The emerging method of femtosecond crystallography (FX) may extend the diffraction resolution accessible from small radiationsensitive 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-angstrom resolution electron densi 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 beta(2)-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.
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