title: Identification and characterization of a linearized B-cell epitope on the pr protein of dengue virus.
abstract: 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.
authors.affiliation: Guangzhou No. 8 People's Hospital, Guangzhou Medical College, Guangzhou 510060, China.|Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Ke-Yu
authors.initials: KY
authors.lastname: Song
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Shi-Hua
authors.initials: SH
authors.lastname: Li
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Li
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Yong-Qiang
authors.initials: YQ
authors.lastname: Deng
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Hong-Jiang
authors.initials: HJ
authors.lastname: Wang
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Ye
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Shun-Ya
authors.initials: SY
authors.lastname: Zhu
authors.affiliation: Department of Microbiology and Immunology, School of Medicine Jinan University, Guangzhou 510632, China.
authors.forename: Zhen-You
authors.initials: ZY
authors.lastname: Jiang
authors.affiliation: Guangzhou No. 8 People's Hospital, Guangzhou Medical College, Guangzhou 510060, China.
authors.forename: Fu-Chun
authors.initials: FC
authors.lastname: Zhang
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: E-De
authors.initials: ED
authors.lastname: Qin
authors.affiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Cheng-Feng
authors.initials: CF
authors.lastname: Qin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D019295:Computational Biology / Q000379:methods; D003715:Dengue / Q000276:immunology / Q000517:prevention & control; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology; D051379:Mice; D016297:Mutagenesis, Site-Directed; D012984:Software; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D018985:Epitopes, B-Lymphocyte; D016056:Immunodominant Epitopes; D014759:Viral Envelope Proteins; D005982:Glutathione Transferase