title: Mutations of an antibody binding energy hot spot on domain III of the dengue 2 envelope glycoprotein exploited for neutralization escape.
abstract: Previous crystallographic studies have identified a total of 11 DENV-2 envelope protein domain III (ED3) residues (K305, F306, K307, V308, V309, K310, I312, Q325, P364, K388, and N390) that interacted, through both side- and main-chain contacts, with the Fab of a dengue virus (DENV) subcomplex-specific neutralizing monoclonal antibody (MAb) 1A1D-2 (Lok et al., 2008). Here, we used DENV-2 recombinant ED3 mutants of the MAb 1A1D-2 structural epitope residues to determine the functional epitope of this MAb. The side-chains of residues K307, K310 and I312 were determined to be functionally critical for MAb binding, and thus constitute a hot spot of binding energy for MAb 1A1D-2 on the DENV-2 ED3. Overall, these findings demonstrate that only a subset of the amino acid residue side-chains within the structural epitope of MAb 1A1D-2 define a functional epitope on the DENV-2 ED3 that is essential for MAb binding and neutralization escape.
authors.affiliation: Department of Pathology, Sealy Center for Vaccine Development, and Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555-0609, USA.
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Gromowski
authors.forename: John T
authors.initials: JT
authors.lastname: Roehrig
authors.forename: Michael S
authors.initials: MS
authors.lastname: Diamond
authors.forename: J Ching
authors.initials: JC
authors.lastname: Lee
authors.forename: Trevor J
authors.initials: TJ
authors.lastname: Pitcher
authors.forename: Alan D T
authors.initials: AD
authors.lastname: Barrett
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D057134:Antibodies, Neutralizing / Q000276:immunology / Q000378:metabolism*; D000914:Antibodies, Viral / Q000276:immunology / Q000378:metabolism; D000915:Antibody Affinity; D002522:Chlorocebus aethiops; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary / Q000235:genetics*; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins