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Title: Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data

Abstract

The X-ray free-electron laser (XFEL) allows the analysis of small weakly diffracting protein crystals, but has required very many crystals to obtain good data. Here we use an XFEL to determine the room temperature atomic structure for the smallest cytoplasmic polyhedrosis virus polyhedra yet characterized, which we failed to solve at a synchrotron. These protein microcrystals, roughly a micron across, accrue within infected cells. We use a new physical model for XFEL diffraction, which better estimates the experimental signal, delivering a high-resolution XFEL structure (1.75 Å), using fewer crystals than previously required for this resolution. The crystal lattice and protein core are conserved compared with a polyhedrin with less than 10% sequence identity. We explain how the conserved biological phenotype, the crystal lattice, is maintained in the face of extreme environmental challenge and massive evolutionary divergence. Our improved methods should open up more challenging biological samples to XFEL analysis.

Authors:
 [1];  [2];  [3];  [1];  [4];  [4];  [4];  [5];  [5];  [4];  [6];  [4];  [5];  [1];  [6]
  1. Univ. of Oxford (United Kingdom). Division of Structural Biology, The Wellcome Trust Centre for Human Genetics.
  2. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  3. Univ. of Oxford (United Kingdom). Division of Structural Biology, The Wellcome Trust Centre for Human Genetics; Yale School of Medicine, New Haven, CT (United States). Molecular Biophysics and Biochemistry
  4. Harwell Science and Innovation Campus, Didcot, Oxfordshire (United Kingdom). Diamond House.
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Physical Biosciences Division.
  6. Univ. of Oxford (United Kingdom). Division of Structural Biology, The Wellcome Trust Centre for Human Genetics; Harwell Science and Innovation Campus, Didcot, Oxfordshire (United Kingdom). Diamond House.
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1190163
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 6; Related Information: Portions of this research were carried out at the Linac Coherent Light Source (LCLS) at the SLAC National Accelerator Laboratory. LCLS is an Office of Science User Facility operated for the U.S. Department of Energy Office of Science by Stanford University.; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
74 ATOMIC AND MOLECULAR PHYSICS; biological sciences; biophysics; virology

Citation Formats

Ginn, Helen M., Messerschmidt, Marc, Ji, Xiaoyun, Zhang, Hanwen, Axford, Danny, Gildea, Richard J., Winter, Graeme, Brewster, Aaron S., Hattne, Johan, Wagner, Armin, Grimes, Jonathan M., Evans, Gwyndaf, Sauter, Nicholas K., Sutton, Geoff, and Stuart, David I.. Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data. United States: N. p., 2015. Web. doi:10.1038/ncomms7435.
Ginn, Helen M., Messerschmidt, Marc, Ji, Xiaoyun, Zhang, Hanwen, Axford, Danny, Gildea, Richard J., Winter, Graeme, Brewster, Aaron S., Hattne, Johan, Wagner, Armin, Grimes, Jonathan M., Evans, Gwyndaf, Sauter, Nicholas K., Sutton, Geoff, & Stuart, David I.. Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data. United States. doi:10.1038/ncomms7435.
Ginn, Helen M., Messerschmidt, Marc, Ji, Xiaoyun, Zhang, Hanwen, Axford, Danny, Gildea, Richard J., Winter, Graeme, Brewster, Aaron S., Hattne, Johan, Wagner, Armin, Grimes, Jonathan M., Evans, Gwyndaf, Sauter, Nicholas K., Sutton, Geoff, and Stuart, David I.. Mon . "Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data". United States. doi:10.1038/ncomms7435. https://www.osti.gov/servlets/purl/1190163.
@article{osti_1190163,
title = {Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data},
author = {Ginn, Helen M. and Messerschmidt, Marc and Ji, Xiaoyun and Zhang, Hanwen and Axford, Danny and Gildea, Richard J. and Winter, Graeme and Brewster, Aaron S. and Hattne, Johan and Wagner, Armin and Grimes, Jonathan M. and Evans, Gwyndaf and Sauter, Nicholas K. and Sutton, Geoff and Stuart, David I.},
abstractNote = {The X-ray free-electron laser (XFEL) allows the analysis of small weakly diffracting protein crystals, but has required very many crystals to obtain good data. Here we use an XFEL to determine the room temperature atomic structure for the smallest cytoplasmic polyhedrosis virus polyhedra yet characterized, which we failed to solve at a synchrotron. These protein microcrystals, roughly a micron across, accrue within infected cells. We use a new physical model for XFEL diffraction, which better estimates the experimental signal, delivering a high-resolution XFEL structure (1.75 Å), using fewer crystals than previously required for this resolution. The crystal lattice and protein core are conserved compared with a polyhedrin with less than 10% sequence identity. We explain how the conserved biological phenotype, the crystal lattice, is maintained in the face of extreme environmental challenge and massive evolutionary divergence. Our improved methods should open up more challenging biological samples to XFEL analysis.},
doi = {10.1038/ncomms7435},
journal = {Nature Communications},
number = ,
volume = 6,
place = {United States},
year = {Mon Mar 09 00:00:00 EDT 2015},
month = {Mon Mar 09 00:00:00 EDT 2015}
}

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