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Serial millisecond crystallography for routine room-temperature structure determination at synchrotrons.

  • Weinert T Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Olieric N Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Cheng R LeadXpro AG, Park InnovAARE, 5234, Villigen PSI, Switzerland.
  • Brünle S Molecular Membrane Biology, Max-Planck Institute of Biophysics, Max-von-Laue-Straße 3, 60438, Frankfurt, Germany.
  • James D Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Ozerov D Science IT, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Gashi D Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Vera L Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Marsh M Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Jaeger K Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Dworkowski F Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Panepucci E Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Basu S Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Skopintsev P Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Doré AS Heptares Therapeutics Ltd, Biopark Broadwater Road, Welwyn Garden City, AL7 3AX, UK.
  • Geng T Heptares Therapeutics Ltd, Biopark Broadwater Road, Welwyn Garden City, AL7 3AX, UK.
  • Cooke RM Heptares Therapeutics Ltd, Biopark Broadwater Road, Welwyn Garden City, AL7 3AX, UK.
  • Liang M Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA, 94025, USA.
  • Prota AE Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Panneels V Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Nogly P Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Ermler U Molecular Membrane Biology, Max-Planck Institute of Biophysics, Max-von-Laue-Straße 3, 60438, Frankfurt, Germany.
  • Schertler G Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Hennig M LeadXpro AG, Park InnovAARE, 5234, Villigen PSI, Switzerland.
  • Steinmetz MO Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Wang M Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
  • Standfuss J Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland. joerg.standfuss@psi.ch.
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  • 2017-09-16
Published in:
  • Nature communications. - 2017
English Historically, room-temperature structure determination was succeeded by cryo-crystallography to mitigate radiation damage. Here, we demonstrate that serial millisecond crystallography at a synchrotron beamline equipped with high-viscosity injector and high frame-rate detector allows typical crystallographic experiments to be performed at room-temperature. Using a crystal scanning approach, we determine the high-resolution structure of the radiation sensitive molybdenum storage protein, demonstrate soaking of the drug colchicine into tubulin and native sulfur phasing of the human G protein-coupled adenosine receptor. Serial crystallographic data for molecular replacement already converges in 1,000-10,000 diffraction patterns, which we collected in 3 to maximally 82 minutes. Compared with serial data we collected at a free-electron laser, the synchrotron data are of slightly lower resolution, however fewer diffraction patterns are needed for de novo phasing. Overall, the data we collected by room-temperature serial crystallography are of comparable quality to cryo-crystallographic data and can be routinely collected at synchrotrons.Serial crystallography was developed for protein crystal data collection with X-ray free-electron lasers. Here the authors present several examples which show that serial crystallography using high-viscosity injectors can also be routinely employed for room-temperature data collection at synchrotrons.
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  • English
Open access status
gold
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https://sonar.ch/global/documents/13443
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