title: Functional analysis of dengue virus (DENV) type 2 envelope protein domain 3 type-specific and DENV complex-reactive critical epitope residues.
abstract: The dengue virus (DENV) envelope protein domain 3 (ED3) is the target of potent virus neutralizing antibodies. The DENV-2 ED3 contains adjacent type-specific and DENV complex-reactive antigenic sites that are composed of a small number of residues that were previously demonstrated to be critical for antibody binding. Site-directed mutagenesis of a DENV-2 16681 infectious clone was used to mutate critical residues in the DENV-2 type-specific (K305A and P384A) and DENV complex-reactive (K310A) antigenic sites. The K305A mutant virus multiplied like the parent virus in mosquito and mammalian cells, as did the P384A mutant virus, which required a compensatory mutation (G330D) for viability. However, the K310A mutant virus could not be recovered. The DENV-2 type-specific critical residue mutations K305A and P384A+G330D reduced the ability of DENV-2 type-specific, but not DENV complex-reactive, mAbs to neutralize virus infectivity and this was directly correlated with mAb binding affinity to the rED3 mutants.
authors.affiliation: Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, Institute for Human Infections and Immunity, and Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0436, USA.
authors.forename: Trevor J
authors.initials: TJ
authors.lastname: Pitcher
authors.affiliation: Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, Institute for Human Infections and Immunity, and Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0436, USA.
authors.forename: Vanessa V
authors.initials: VV
authors.lastname: Sarathy
authors.affiliation: Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, Institute for Human Infections and Immunity, and Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0436, USA.
authors.forename: Kiyohiko
authors.initials: K
authors.lastname: Matsui
authors.affiliation: Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, Institute for Human Infections and Immunity, and Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0436, USA.
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Gromowski
authors.affiliation: Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Public Health Service, US Department of Health and Human Services, Fort Collins, CO 80521, USA.
authors.forename: Claire Y-H
authors.initials: CY
authors.lastname: Huang
authors.affiliation: Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, Institute for Human Infections and Immunity, and Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0436, USA.
authors.forename: Alan D T
authors.initials: ADT
authors.lastname: Barrett
mesh_terms: D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D004252:DNA Mutational Analysis; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D050296:Microbial Viability; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D014779:Virus Replication
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins