Identification and characterization of a linearized B-cell epitope on the pr protein of dengue virus
| Autor: | Ke-Yu Song, Fuchun Zhang, Cheng-Feng Qin, E-De Qin, Qing Ye, Hong-Jiang Wang, Hui Zhao, Shihua Li, Yong-Qiang Deng, Zhen-You Jiang, Xiaofeng Li, Shun-Ya Zhu |
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| Rok vydání: | 2013 |
| Předmět: |
viruses
Biology Dengue virus medicine.disease_cause Antibodies Viral Epitope Dengue Mice Viral Envelope Proteins Virology medicine Animals Humans Amino Acid Sequence Peptide sequence Glutathione Transferase Antiserum Linear epitope Immunodominant Epitopes virus diseases Antibodies Monoclonal Computational Biology Dengue Virus Epitope mapping Virion assembly biology.protein Mutagenesis Site-Directed Epitopes B-Lymphocyte Antibody Epitope Mapping Software |
| Zdroj: | The Journal of general virology. 94(Pt 7) |
| ISSN: | 1465-2099 |
| Popis: | The four serotypes of dengue virus (DENV) represent one of the major mosquito-borne pathogens globally; so far no vaccine or specific antiviral is available. During virion maturation, the pr protein is cleaved from its precursor form the prM protein on the surface of immature DENV by host protease. Recent findings have demonstrated that the pr protein not only played critical roles in virion assembly and maturation, but was also involved in antibody-dependent enhancement of DENV infection. However, the B-cell epitopes on the pr protein of DENV have not been well characterized. In this study, a set of 11 partially overlapping peptides spanning the entire pr protein of DENV-2 were fused with glutathione S-transferase and expressed in Escherichia coli. ELISA screening with murine hyperimmune antiserum against immature DENV identified the P8 peptide (57KQNEPEDIDCWCNST71) in the pr protein as the major immunodominant epitope. Fine mapping by truncated protein assays confirmed the 8-e peptide 57KQNEPEDI64 was the smallest unit capable of antibody binding. Importantly, the 8-e epitope reacted with sera from dengue fever patients. Site-directed mutagenesis revealed the asparagine residue at position 59 was important for epitope recognition. The 8-e epitope coincided well with the B-cell epitopes predicted by Immune Epitope Database analysis, and 3D structural modelling mapped the 8-e peptide on the surface of prM-E heterodimers. Overall, our findings characterized a linearized B-cell epitope on the pr protein of DENV, which will help to understand the life cycle of DENV and pathogenesis of dengue infections in human. |
| Databáze: | OpenAIRE |
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