Structure of CPV17 polyhedrin determined by the improved analysis of serial femtosecond crystallographic data.
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.
Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK.
Helen M
HM
Ginn
SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
Marc
M
Messerschmidt
1] Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK [2] Molecular Biophysics and Biochemistry, Yale School of Medicine, 333 Cedar Street, New Haven, Connecticut 06510, USA.
Xiaoyun
X
Ji
Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK.
Hanwen
H
Zhang
Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
Danny
D
Axford
Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
Richard J
0000000150386958
RJ
Gildea
Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
Graeme
G
Winter
Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, USA.
Aaron S
AS
Brewster
Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, USA.
Johan
0000000289360912
J
Hattne
Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
Armin
A
Wagner
1] Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK [2] Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
Jonathan M
JM
Grimes
Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
Gwyndaf
0000000260792201
G
Evans
Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, USA.
Nicholas K
NK
Sauter
Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK.
Geoff
G
Sutton
1] Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK [2] Diamond House, Diamond Light Source, Harwell Science &Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
David I
DI
Stuart
D000818:Animals; D003461:Crystallography / Q000379:methods*; D009033:Culicidae / Q000821:virology*; D005822:Genetic Vectors / Q000235:genetics; D007091:Image Processing, Computer-Assisted; D008958:Models, Molecular; D011487:Protein Conformation; D012087:Reoviridae / Q000737:chemistry*; D015678:Viral Structural Proteins / Q000737:chemistry*
D015678:Viral Structural Proteins