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

Helen M. Ginn, Marc Messerschmidt, Xiaoyun Ji, Hanwen Zhang, Danny Axford, Richard J. Gildea, Graeme Winter, Aaron S. Brewster, Johan Hattne, Armin Wagner, Jonathan M. Grimes, Gwyndaf Evans, Nicholas K. Sauter, Geoff Sutton, David I. Stuart

Research output: Contribution to journalArticle

43 Citations (Scopus)

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 (1.75â €‰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.

Original languageEnglish (US)
Article number6435
JournalNature Communications
Volume6
DOIs
StatePublished - Mar 9 2015
Externally publishedYes

Fingerprint

X ray lasers
Free electron lasers
free electron lasers
Lasers
X-Rays
Electrons
proteins
x rays
crystal lattices
Crystal lattices
Crystals
Reoviridae
crystals
Microcrystals
Synchrotrons
Proteins
phenotype
microcrystals
viruses
polyhedrons

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data. / 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.

In: Nature Communications, Vol. 6, 6435, 09.03.2015.

Research output: Contribution to journalArticle

Ginn, HM, Messerschmidt, M, Ji, X, Zhang, H, Axford, D, Gildea, RJ, Winter, G, Brewster, AS, Hattne, J, Wagner, A, Grimes, JM, Evans, G, Sauter, NK, Sutton, G & Stuart, DI 2015, 'Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data', Nature Communications, vol. 6, 6435. https://doi.org/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. In: Nature Communications. 2015 ; Vol. 6.
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