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Modeling the structure of mAb 14B7 bound to the anthrax protective antigen
Author(s) -
Sivasubramanian Arvind,
Maynard Jennifer A.,
Gray Jeffrey J.
Publication year - 2008
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.21595
Subject(s) - docking (animal) , homology modeling , monoclonal antibody , dock , epitope , binding site , chemistry , antigen , protein data bank (rcsb pdb) , antibody , macromolecular docking , anthrax toxin , molecular model , computational biology , stereochemistry , biology , biochemistry , genetics , enzyme , recombinant dna , fusion protein , gene , medicine , nursing
Abstract The anthrax protective antigen (PA) is a key component of the tripartite anthrax toxin. Monoclonal antibody (mAb) 14B7 and its engineered, affinity‐matured variants have been shown to be effective in blocking PA binding to cellular receptors and mitigating anthrax toxicity. Here, we perform computational structural modeling of the mAb 14B7‐PA interaction. Our objectives are to determine the structure of the 14B7‐PA complex, to deduce a structural explanation for the affinity maturation from the docking models, and to study the effect of inaccuracies in the antibody homology model on docking. We used the RosettaDock program to dock PA with the mAb 14B7 crystal structure or homology model. Our simulations generate two distinct binding orientations consistent with experimental residue mutations that diminish 14B7‐PA binding. Furthermore, the models suggest new site‐directed mutations to positively identify one of these two solutions as the correct 14B7‐PA docking orientation. The models indicate that PA regions 648–660 and 712–720 may be important for 14B7 binding in addition to the known PA epitope, and the binding interfaces are similar to that seen in the PA complex with cellular receptor CMG2. Antibody residues involved in affinity maturation do not contact the antigen in the docking models, suggesting that affinity maturation in the 14B7 family does not result from direct enhancements of antibody–antigen contacts. Docking the homology model produces low‐resolution representations of the crystal structure docking orientations, but homology model docking is frustrated by antibody H3 loop conformation errors. This work demonstrates the usefulness and limitations of computational structure prediction for the development of antibody therapeutics, and reemphasizes the need for flexible backbone docking algorithms to achieve high‐resolution docking using homology models. Proteins 2008. © 2007 Wiley‐Liss, Inc.