Structural and functional characterization of an agonistic anti-human EphA2 monoclonal antibody

Li Peng; Vaheh Oganesyan; Melissa M Damschroder; Herren Wu; William F Dall'Acqua

doi:10.1016/j.jmb.2011.08.018

Structural and functional characterization of an agonistic anti-human EphA2 monoclonal antibody

J Mol Biol. 2011 Oct 21;413(2):390-405. doi: 10.1016/j.jmb.2011.08.018. Epub 2011 Aug 16.

Authors

Li Peng¹, Vaheh Oganesyan, Melissa M Damschroder, Herren Wu, William F Dall'Acqua

Affiliation

¹ Department of Antibody Discovery and Protein Engineering, MedImmune, Gaithersburg, MD 20878, USA.

PMID: 21867711
DOI: 10.1016/j.jmb.2011.08.018

Abstract

We report here the three-dimensional structure of human ephrin type A receptor 2 (EphA2) bound to the Fab (fragment antigen binding) of an agonistic human antibody (1C1; IgG1/κ). The structure of the corresponding complex was solved at a resolution of 2.5 Å using molecular replacement and constitutes the first reported structure of a human ephrin receptor bound to an antibody. We have also defined the corresponding functional epitope using a mutagenesis-based approach. This study revealed discrete structural features that determine the fine specificity of 1C1 to EphA2. Our data also provided a molecular basis for 1C1 mechanism of action. More precisely, we propose that its agonistic, internalizing properties are the result of ligand mimicry by the third heavy-chain complementarity-determining region of 1C1. Because EphA2 is an important contributor to cancer formation and progression, these findings may have implications for designing the next generation of anti-tumor therapies.

MeSH terms

Amino Acid Sequence
Animals
Antibodies, Anti-Idiotypic / chemistry*
Antibodies, Anti-Idiotypic / immunology
Antibodies, Anti-Idiotypic / metabolism
Antibodies, Monoclonal / chemistry*
Antibodies, Monoclonal / immunology
Antibodies, Monoclonal / metabolism
Crystallography, X-Ray
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Humans
Immunoglobulin Fab Fragments / chemistry*
Immunoglobulin Fab Fragments / immunology
Immunoglobulin Fab Fragments / metabolism
Immunoglobulin G / chemistry*
Immunoglobulin G / immunology
Immunoglobulin G / metabolism
Immunoglobulin kappa-Chains / chemistry*
Immunoglobulin kappa-Chains / immunology
Immunoglobulin kappa-Chains / metabolism
Male
Mice
Models, Chemical
Molecular Sequence Data
Phosphorylation
Prostatic Neoplasms / metabolism
Protein Structure, Secondary
Receptor, EphA2 / agonists*
Receptor, EphA2 / immunology
Receptor, EphA2 / metabolism
Sequence Homology, Amino Acid
Tumor Cells, Cultured

Substances

Antibodies, Anti-Idiotypic
Antibodies, Monoclonal
Immunoglobulin Fab Fragments
Immunoglobulin G
Immunoglobulin kappa-Chains
Receptor, EphA2

Associated data

PDB/3SKJ