Format

Send to

Choose Destination
Nat Commun. 2017 Sep 14;8(1):542. doi: 10.1038/s41467-017-00630-4.

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

Author information

1
Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
2
LeadXpro AG, Park InnovAARE, 5234, Villigen PSI, Switzerland.
3
Molecular Membrane Biology, Max-Planck Institute of Biophysics, Max-von-Laue-Straße 3, 60438, Frankfurt, Germany.
4
Science IT, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
5
SwissFEL, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
6
Macromolecular Crystallography, Swiss Light Source, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland.
7
Heptares Therapeutics Ltd, Biopark Broadwater Road, Welwyn Garden City, AL7 3AX, UK.
8
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA, 94025, USA.
9
Department of Biology, ETH Zurich, 8093, Zürich, Switzerland.
10
University of Basel, Biozentrum, Basel, 4056, Switzerland.
11
Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232, Villigen PSI, Switzerland. joerg.standfuss@psi.ch.

Abstract

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.

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center