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Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus

Porcine deltacoronavirus (PDCoV), first identified in 2012, is a swine enteropathogen now found in many countries. The nucleocapsid (N) protein, a core component of PDCoV, is essential for virus replication and is a significant candidate in the development of diagnostics for PDCoV. In this study, mo...

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Autores principales: Fu, Jiayu, Chen, Rui, Hu, Jingfei, Qu, Huan, Zhao, Yujia, Cao, Sanjie, Wen, Xintian, Wen, Yiping, Wu, Rui, Zhao, Qin, Ma, Xiaoping, Huang, Xiaobo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013544/
https://www.ncbi.nlm.nih.gov/pubmed/31963776
http://dx.doi.org/10.3390/ijms21020648
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author Fu, Jiayu
Chen, Rui
Hu, Jingfei
Qu, Huan
Zhao, Yujia
Cao, Sanjie
Wen, Xintian
Wen, Yiping
Wu, Rui
Zhao, Qin
Ma, Xiaoping
Huang, Xiaobo
author_facet Fu, Jiayu
Chen, Rui
Hu, Jingfei
Qu, Huan
Zhao, Yujia
Cao, Sanjie
Wen, Xintian
Wen, Yiping
Wu, Rui
Zhao, Qin
Ma, Xiaoping
Huang, Xiaobo
author_sort Fu, Jiayu
collection PubMed
description Porcine deltacoronavirus (PDCoV), first identified in 2012, is a swine enteropathogen now found in many countries. The nucleocapsid (N) protein, a core component of PDCoV, is essential for virus replication and is a significant candidate in the development of diagnostics for PDCoV. In this study, monoclonal antibodies (mAbs) were generated and tested for reactivity with three truncations of the full protein (N1, N2, N3) that contained partial overlaps; of the five monoclonals chosen tested, each reacted with only the N3 truncation. The antibody designated 4E88 had highest binding affinity with the N protein and was chosen for in-depth examination. The 4E88 epitope was located to amino acids 308-AKPKQQKKPKK-318 by testing the 4E88 monoclonal for reactivity with a series of N3 truncations, then the minimal epitope, 309-KPKQQKKPK-317 (designated EP-4E88), was pinpointed by testing the 4E88 monoclonal for reactivity with a series of synthetic peptides of this region. Homology analysis showed that the EP-4E88 sequence is highly conserved among PDCoV strains, and also shares high similarity with sparrow coronavirus (HKU17), Asian leopard cat coronavirus (ALCCoV), quail coronavirus (UAE-HKU30), and sparrow deltacoronavirus (SpDCoV). Of note, the PDCoV EP-4E88 sequence shared very low similarity (<22.2%) with other porcine coronaviruses (PEDV, TGEV, PRCV, SADS-CoV, PHEV), demonstrating that it is an epitope that can be used for distinguishing PDCoV and other porcine coronavirus. 3D structural analysis revealed that amino acids of EP-4E88 were in close proximity and may be exposed on the surface of the N protein.
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spelling pubmed-70135442020-03-09 Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus Fu, Jiayu Chen, Rui Hu, Jingfei Qu, Huan Zhao, Yujia Cao, Sanjie Wen, Xintian Wen, Yiping Wu, Rui Zhao, Qin Ma, Xiaoping Huang, Xiaobo Int J Mol Sci Article Porcine deltacoronavirus (PDCoV), first identified in 2012, is a swine enteropathogen now found in many countries. The nucleocapsid (N) protein, a core component of PDCoV, is essential for virus replication and is a significant candidate in the development of diagnostics for PDCoV. In this study, monoclonal antibodies (mAbs) were generated and tested for reactivity with three truncations of the full protein (N1, N2, N3) that contained partial overlaps; of the five monoclonals chosen tested, each reacted with only the N3 truncation. The antibody designated 4E88 had highest binding affinity with the N protein and was chosen for in-depth examination. The 4E88 epitope was located to amino acids 308-AKPKQQKKPKK-318 by testing the 4E88 monoclonal for reactivity with a series of N3 truncations, then the minimal epitope, 309-KPKQQKKPK-317 (designated EP-4E88), was pinpointed by testing the 4E88 monoclonal for reactivity with a series of synthetic peptides of this region. Homology analysis showed that the EP-4E88 sequence is highly conserved among PDCoV strains, and also shares high similarity with sparrow coronavirus (HKU17), Asian leopard cat coronavirus (ALCCoV), quail coronavirus (UAE-HKU30), and sparrow deltacoronavirus (SpDCoV). Of note, the PDCoV EP-4E88 sequence shared very low similarity (<22.2%) with other porcine coronaviruses (PEDV, TGEV, PRCV, SADS-CoV, PHEV), demonstrating that it is an epitope that can be used for distinguishing PDCoV and other porcine coronavirus. 3D structural analysis revealed that amino acids of EP-4E88 were in close proximity and may be exposed on the surface of the N protein. MDPI 2020-01-19 /pmc/articles/PMC7013544/ /pubmed/31963776 http://dx.doi.org/10.3390/ijms21020648 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fu, Jiayu
Chen, Rui
Hu, Jingfei
Qu, Huan
Zhao, Yujia
Cao, Sanjie
Wen, Xintian
Wen, Yiping
Wu, Rui
Zhao, Qin
Ma, Xiaoping
Huang, Xiaobo
Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus
title Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus
title_full Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus
title_fullStr Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus
title_full_unstemmed Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus
title_short Identification of a Novel Linear B-Cell Epitope on the Nucleocapsid Protein of Porcine Deltacoronavirus
title_sort identification of a novel linear b-cell epitope on the nucleocapsid protein of porcine deltacoronavirus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013544/
https://www.ncbi.nlm.nih.gov/pubmed/31963776
http://dx.doi.org/10.3390/ijms21020648
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