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

doi:10.1038/ncomms7435

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 journal › Article › peer-review

53 Scopus citations

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 language	English (US)
Article number	6435
Journal	Nature communications
Volume	6
DOIs	https://doi.org/10.1038/ncomms7435
State	Published - Mar 9 2015
Externally published	Yes

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

Access to Document

10.1038/ncomms7435

Cite this

Ginn, H. M., Messerschmidt, M., Ji, X., Zhang, H., Axford, D., Gildea, R. J., Winter, G., Brewster, A. S., Hattne, J., Wagner, A., Grimes, J. M., Evans, G., Sauter, N. K., Sutton, G., & Stuart, D. I. (2015). Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data. Nature communications, 6, Article 6435. https://doi.org/10.1038/ncomms7435

@article{41ff7234be0940a7bdd60aef5152a6a7,

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{\^a} €‰{\AA}), 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.",

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

year = "2015",

month = mar,

day = "9",

doi = "10.1038/ncomms7435",

language = "English (US)",

volume = "6",

journal = "Nature communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data

AU - Ginn, Helen M.

AU - Messerschmidt, Marc

AU - Ji, Xiaoyun

AU - Zhang, Hanwen

AU - Axford, Danny

AU - Gildea, Richard J.

AU - Winter, Graeme

AU - Brewster, Aaron S.

AU - Hattne, Johan

AU - Wagner, Armin

AU - Grimes, Jonathan M.

AU - Evans, Gwyndaf

AU - Sauter, Nicholas K.

AU - Sutton, Geoff

AU - Stuart, David I.

PY - 2015/3/9

Y1 - 2015/3/9

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84924386737&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84924386737&partnerID=8YFLogxK

U2 - 10.1038/ncomms7435

DO - 10.1038/ncomms7435

M3 - Article

C2 - 25751308

AN - SCOPUS:84924386737

SN - 2041-1723

VL - 6

JO - Nature communications

JF - Nature communications

M1 - 6435

ER -

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

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this