Database of articles cited by EMDB/PDB/SASBDB data

Keywords
Structure methods	All X-ray diffraction NMR electron microscopy small angle scattering neutron diffraction fiber diffraction powder diffraction infrared spectroscopy EPR fluorescence transfer theoretical model Hybrid
Author
Journal
IF	>30 >20 >10 >5 all

Title	Structural roles of PCV2 capsid protein N-terminus in PCV2 particle assembly and identification of PCV2 type-specific neutralizing epitope.
Journal, issue, pages	PLoS Pathog, Vol. 15, Issue 3, Page e1007562, Year 2019
Publish date	Mar 1, 2019
Authors	Xiaobing Mo / Xiangdong Li / Bo Yin / Junhua Deng / Kegong Tian / Adam Yuan /
PubMed Abstract	Postweaning multisystemic wasting disease (PMWS) in piglets caused by porcine circovirus type 2 (PCV2) is one of the major threats to most pig farms worldwide. Among all the PCV types, PCV2 is the ...Postweaning multisystemic wasting disease (PMWS) in piglets caused by porcine circovirus type 2 (PCV2) is one of the major threats to most pig farms worldwide. Among all the PCV types, PCV2 is the dominant genotype causing PMWS and associated diseases. Considerable efforts were made to study the virus-like-particle (VLP) assembly and the specific PCV2-associated epitope(s) in order to establish the solid foundation for engineered PCV2 vaccine development. Although the N-terminal fragment including Nuclear Localization Signal (NLS) sequence seems important for recombinant PCV2 capsid protein expression and VLP assembly, the detailed structural and functional information regarding this important fragment are largely unknown. In this study, we report crystal structure of PCV2 VLP assembled from N-terminal NLS truncated PCV2 capsid protein at 2.8 Å resolution and cryo-EM structure of PCV2 VLP assembled from full-length PCV2 capsid protein at 4.1Å resolution. Our in vitro PCV2 VLP assembly results show that NLS-truncated PCV2 capsid protein only forms instable VLPs which were easily disassembled in solution, whereas full-length PCV2 capsid protein forms stable VLPs due to interaction between 15PRSHLGQILRRRP27 (α-helix) and 33RHRYRWRRKN42 (NLS-B) in a repeated manner. In addition, our results also showed that N-terminal truncation of PCV2 capsid protein up to 27 residues still forms PCV2 particles in solution with similar size and immunogenicity, while N-terminal truncation of PCV2 capsid protein with more than 30 residues is not able to form stable PCV2 particles in solution, demonstrating the importance of interaction between the α-helix at N-terminal and NLS-B in PCV2 VLP formation. Moreover, we also report the cryo-EM structure of PCV2 VLP in complex with 3H11-Fab, a PCV2 type-specific neutralizing antibody, at 15 Å resolution. MAb-3H11 specifically recognizes one exposed epitope located on the VLP surface EF-loop (residues 128-143), which is further confirmed by PCV1-PCV2 epitope swapping assay. Hence, our results have revealed the structural roles of N-terminal fragment of PCV2 capsid protein in PCV2 particle assembly and pinpointed one PCV2 type-specific neutralizing epitope for the first time, which could provide clear clue for next generation PCV2 vaccine and diagnostic kits development.
External links	PLoS Pathog / PubMed:30822338 / PubMed Central
Methods	EM (single particle) / X-ray diffraction
Resolution	2.8 - 15.0 Å
Structure data	6746 5zbo EMDB-6746: CryoEM structure of PCV2 VLPs PDB-5zbo: Cryo-EM structure of PCV2 VLPs Method: EM (single particle) / Resolution: 4.12 Å EMDB-6961: Cryo-EM structure of PCV2 VLP+Fab fragments of mAb-3H11 Method: EM (single particle) / Resolution: 15.0 Å PDB-5zju: Crystal structure of in vitro expressed and assembled PCV2 Virus-like Particle Method: X-RAY DIFFRACTION / Resolution: 2.8 Å
Chemicals	ChemComp-HOH: WATER
Source	porcine circovirus 2
Keywords	DNA BINDING PROTEIN / Porcine circle virus type 2 (PCV2) / capsid protein / virus-like-particle (VLP) / type-specific epitope / VIRUS / PCV2 capsid protein