Cargando…

Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle

Betanodavirus infection causes fatal disease of viral nervous necrosis in many cultured marine and freshwater fish worldwide and the virus-like particles (VLP) are effective vaccines against betanodavirus. But vaccine and viral vector designs of betanodavirus VLP based on their structures remain lac...

Descripción completa

Detalles Bibliográficos
Autores principales: Xie, Junfeng, Li, Kunpeng, Gao, Yuanzhu, Huang, Runqing, Lai, Yuxiong, Shi, Yan, Yang, Shaowei, Zhu, Guohua, Zhang, Qinfen, He, Jianguo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710043/
https://www.ncbi.nlm.nih.gov/pubmed/26754256
http://dx.doi.org/10.1186/s13567-015-0294-9
_version_ 1782409767593967616
author Xie, Junfeng
Li, Kunpeng
Gao, Yuanzhu
Huang, Runqing
Lai, Yuxiong
Shi, Yan
Yang, Shaowei
Zhu, Guohua
Zhang, Qinfen
He, Jianguo
author_facet Xie, Junfeng
Li, Kunpeng
Gao, Yuanzhu
Huang, Runqing
Lai, Yuxiong
Shi, Yan
Yang, Shaowei
Zhu, Guohua
Zhang, Qinfen
He, Jianguo
author_sort Xie, Junfeng
collection PubMed
description Betanodavirus infection causes fatal disease of viral nervous necrosis in many cultured marine and freshwater fish worldwide and the virus-like particles (VLP) are effective vaccines against betanodavirus. But vaccine and viral vector designs of betanodavirus VLP based on their structures remain lacking. Here, the three-dimensional structure of orange-spotted grouper nervous necrosis virus (OGNNV) VLP (RBS) at 3.9 Å reveals the organization of capsid proteins (CP). Based on the structural results, seven putative important sites were selected to genetically insert a 6× histidine (His)-tag for VLP formation screen, resulting in four His-tagged VLP (HV) at positions N-terminus, Ala220, Pro292 and C-terminus. The His-tags of N-terminal HV (NHV) were concealed inside virions while those of 220HV and C-terminal HV (CHV) were displayed at the outer surface. NHV, 220HV and CHV maintained the same cell entry ability as RBS in the Asian sea bass (SB) cell line, indicating that their similar surface structures can be recognized by the cellular entry receptor(s). For application of vaccine design, chromatography-purified CHV could provoke NNV-specific antibody responses as strong as those of RBS in a sea bass immunization assay. Furthermore, in carrying capacity assays, N-terminus and Ala220 can only carry short peptides and C-terminus can even accommodate large protein such as GFP to generate fluorescent VLP (CGV). For application of a viral vector, CGV could be real-time visualized to enter SB cells in invasion study. All the results confirmed that the C-terminus of CP is a suitable site to accommodate foreign peptides for vaccine design and viral vector development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13567-015-0294-9) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4710043
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-47100432016-01-13 Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle Xie, Junfeng Li, Kunpeng Gao, Yuanzhu Huang, Runqing Lai, Yuxiong Shi, Yan Yang, Shaowei Zhu, Guohua Zhang, Qinfen He, Jianguo Vet Res Research Article Betanodavirus infection causes fatal disease of viral nervous necrosis in many cultured marine and freshwater fish worldwide and the virus-like particles (VLP) are effective vaccines against betanodavirus. But vaccine and viral vector designs of betanodavirus VLP based on their structures remain lacking. Here, the three-dimensional structure of orange-spotted grouper nervous necrosis virus (OGNNV) VLP (RBS) at 3.9 Å reveals the organization of capsid proteins (CP). Based on the structural results, seven putative important sites were selected to genetically insert a 6× histidine (His)-tag for VLP formation screen, resulting in four His-tagged VLP (HV) at positions N-terminus, Ala220, Pro292 and C-terminus. The His-tags of N-terminal HV (NHV) were concealed inside virions while those of 220HV and C-terminal HV (CHV) were displayed at the outer surface. NHV, 220HV and CHV maintained the same cell entry ability as RBS in the Asian sea bass (SB) cell line, indicating that their similar surface structures can be recognized by the cellular entry receptor(s). For application of vaccine design, chromatography-purified CHV could provoke NNV-specific antibody responses as strong as those of RBS in a sea bass immunization assay. Furthermore, in carrying capacity assays, N-terminus and Ala220 can only carry short peptides and C-terminus can even accommodate large protein such as GFP to generate fluorescent VLP (CGV). For application of a viral vector, CGV could be real-time visualized to enter SB cells in invasion study. All the results confirmed that the C-terminus of CP is a suitable site to accommodate foreign peptides for vaccine design and viral vector development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13567-015-0294-9) contains supplementary material, which is available to authorized users. BioMed Central 2016-01-11 2016 /pmc/articles/PMC4710043/ /pubmed/26754256 http://dx.doi.org/10.1186/s13567-015-0294-9 Text en © Xie et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Xie, Junfeng
Li, Kunpeng
Gao, Yuanzhu
Huang, Runqing
Lai, Yuxiong
Shi, Yan
Yang, Shaowei
Zhu, Guohua
Zhang, Qinfen
He, Jianguo
Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
title Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
title_full Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
title_fullStr Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
title_full_unstemmed Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
title_short Structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
title_sort structural analysis and insertion study reveal the ideal sites for surface displaying foreign peptides on a betanodavirus-like particle
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710043/
https://www.ncbi.nlm.nih.gov/pubmed/26754256
http://dx.doi.org/10.1186/s13567-015-0294-9
work_keys_str_mv AT xiejunfeng structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT likunpeng structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT gaoyuanzhu structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT huangrunqing structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT laiyuxiong structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT shiyan structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT yangshaowei structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT zhuguohua structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT zhangqinfen structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle
AT hejianguo structuralanalysisandinsertionstudyrevealtheidealsitesforsurfacedisplayingforeignpeptidesonabetanodaviruslikeparticle