Journal
IUCRJ
Volume 5, Issue -, Pages 727-736Publisher
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S205225251801120X
Keywords
single-particle imaging; three-dimensional virus reconstructions; XFELs
Funding
- US Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-76SF00515]
- Helmholtz Association's Initiative and Networking Fund
- Russian Science Foundation [18-41-06001]
- Joachim Herz Foundation Hamburg
- Program-Oriented Funds of the Helmholtz Association
- Swedish Research Council
- Swedish Foundation for Strategic Research
- US National Science Foundation (NSF) Science and Technology Center BioXFEL Award [1231306]
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The analysis of a single-particle imaging (SPI) experiment performed at the AMO beamline at LCLS as part of the SPI initiative is presented here. A workflow for the three-dimensional virus reconstruction of the PR772 bacteriophage from measured single-particle data is developed. It consists of several well defined steps including single-hit diffraction data classification, refined filtering of the classified data, reconstruction of three-dimensional scattered intensity from the experimental diffraction patterns by orientation determination and a final three-dimensional reconstruction of the virus electron density without symmetry constraints. The analysis developed here revealed and quantified nanoscale features of the PR772 virus measured in this experiment, with the obtained resolution better than 10 nm, with a clear indication that the structure was compressed in one direction and, as such, deviates from ideal icosahedral symmetry.
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