Computation-guided backbone grafting of a discontinuous motif onto a protein scaffold.

Publication Type:

Journal Article

Source:

Science (New York, N.Y.), Volume 334, Issue 6054, p.373-6 (2011)

Keywords:

2011, Algorithms, Amino Acid Motifs, Amino Acid Sequence, Antibodies, Monoclonal, Antibodies, Neutralizing, Antibody Affinity, Antibody Specificity, Antigens, CD4, Basic Sciences Division, Center-Authored Paper, Computational Biology, Computer Simulation, Crystallography, X-Ray, Epitopes, HIV Antibodies, HIV Envelope Protein gp120, Models, Molecular, Molecular Mimicry, Molecular Sequence Data, mutagenesis, November 2011, Protein Conformation, Protein Engineering, Protein Interaction Domains and Motifs, Surface Plasmon Resonance

Abstract:

The manipulation of protein backbone structure to control interaction and function is a challenge for protein engineering. We integrated computational design with experimental selection for grafting the backbone and side chains of a two-segment HIV gp120 epitope, targeted by the cross-neutralizing antibody b12, onto an unrelated scaffold protein. The final scaffolds bound b12 with high specificity and with affinity similar to that of gp120, and crystallographic analysis of a scaffold bound to b12 revealed high structural mimicry of the gp120-b12 complex structure. The method can be generalized to design other functional proteins through backbone grafting.