Advances in X-ray free electron laser (XFEL) diffraction data processing applied to the crystal structure of the synaptotagmin-1 / SNARE complex

X-ray free electron lasers (XFELs) reduce the effects of radiation damage on macromolecular diffraction data and thereby extend the limiting resolution. Previously, we adapted classical post-refinement techniques to XFEL diffraction data to produce accurate diffraction data sets from a limited numbe...

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Bibliographic Details
Published in:eLife 2016-10, Vol.5 (2016)
Main Authors: Lyubimov, Artem Y, Uervirojnangkoorn, Monarin, Zeldin, Oliver B, Zhou, Qiangjun, Zhao, Minglei, Brewster, Aaron S, Michels-Clark, Tara, Holton, James M, Sauter, Nicholas K, Weis, William I, Brunger, Axel T
Format: Article
Language:English
Subjects:
Biophysics
Biophysics and Structural Biology
Crystal lattices
Crystal structure
Crystallization
Crystals
Data processing
Datasets
Diffraction
Image Processing, Computer-Assisted - methods
Laboratories
Lasers
macromolecular crystallography
Macromolecules
Medical research
Microscopy, Electron, Transmission - methods
post-refinement
radiation damage
Research Advance
SNAP receptors
SNARE Proteins - chemistry
Synaptotagmin
Synaptotagmin I - chemistry
X-ray free electron laser
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Summary:X-ray free electron lasers (XFELs) reduce the effects of radiation damage on macromolecular diffraction data and thereby extend the limiting resolution. Previously, we adapted classical post-refinement techniques to XFEL diffraction data to produce accurate diffraction data sets from a limited number of diffraction images (Uervirojnangkoorn et al., 2015), and went on to use these techniques to obtain a complete data set from crystals of the synaptotagmin-1 / SNARE complex and to determine the structure at 3.5 Å resolution (Zhou et al., 2015). Here, we describe new advances in our methods and present a reprocessed XFEL data set of the synaptotagmin-1 / SNARE complex. The reprocessing produced small improvements in electron density maps and the refined atomic model. The maps also contained more information than those of a lower resolution (4.1 Å) synchrotron data set. Processing a set of simulated XFEL diffraction images revealed that our methods yield accurate data and atomic models.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.18740