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High-speed fixed-target serial virus crystallography.

  • Roedig P Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
  • Ginn HM Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Pakendorf T Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
  • Sutton G Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Harlos K Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Walter TS Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Meyer J Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
  • Fischer P Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
  • Duman R Diamond Light Source Limited, Didcot, UK.
  • Vartiainen I Institute of Photonics, University of Eastern Finland, Joensuu, Finland.
  • Reime B Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
  • Warmer M Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
  • Brewster AS Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
  • Young ID Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
  • Michels-Clark T Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
  • Sauter NK Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
  • Kotecha A Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Kelly J School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK.
  • Rowlands DJ School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK.
  • Sikorsky M Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
  • Nelson S Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
  • Damiani DS Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
  • Alonso-Mori R Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
  • Ren J Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Fry EE Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • David C Paul Scherrer Institut, Villigen-PSI, Switzerland.
  • Stuart DI Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK.
  • Wagner A Diamond Light Source Limited, Didcot, UK.
  • Meents A Photon Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
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  • 2017-06-20
Published in:
  • Nature methods. - 2017
Algorithms
Crystallography, X-Ray
Image Enhancement
Image Interpretation, Computer-Assisted
Imaging, Three-Dimensional
Reproducibility of Results
Sample Size
Sensitivity and Specificity
Viruses
English We report a method for serial X-ray crystallography at X-ray free-electron lasers (XFELs), which allows for full use of the current 120-Hz repetition rate of the Linear Coherent Light Source (LCLS). Using a micropatterned silicon chip in combination with the high-speed Roadrunner goniometer for sample delivery, we were able to determine the crystal structures of the picornavirus bovine enterovirus 2 (BEV2) and the cytoplasmic polyhedrosis virus type 18 polyhedrin, with total data collection times of less than 14 and 10 min, respectively. Our method requires only micrograms of sample and should therefore broaden the applicability of serial femtosecond crystallography to challenging projects for which only limited sample amounts are available. By synchronizing the sample exchange to the XFEL repetition rate, our method allows for most efficient use of the limited beam time available at XFELs and should enable a substantial increase in sample throughput at these facilities.
Language
  • English
Open access status
green
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Persistent URL
https://sonar.ch/global/documents/44811
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