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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Algorithms
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Amino Acid Motifs
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Amino Acid Sequence
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / immunology
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Antibodies, Monoclonal / metabolism
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Antibodies, Neutralizing / chemistry*
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Antibodies, Neutralizing / immunology*
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Antibodies, Neutralizing / metabolism
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Antibody Affinity
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Antibody Specificity
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CD4 Antigens / metabolism
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Computational Biology
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Computer Simulation
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Crystallography, X-Ray
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Epitopes / immunology
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HIV Antibodies / chemistry
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HIV Antibodies / immunology*
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HIV Antibodies / metabolism
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HIV Envelope Protein gp120 / chemistry*
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HIV Envelope Protein gp120 / immunology*
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HIV Envelope Protein gp120 / metabolism
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Models, Molecular
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Molecular Mimicry
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Molecular Sequence Data
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Mutagenesis
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Protein Conformation
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Protein Engineering*
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Protein Interaction Domains and Motifs
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Surface Plasmon Resonance
Substances
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Antibodies, Monoclonal
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Antibodies, Neutralizing
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CD4 Antigens
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Epitopes
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HIV Antibodies
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HIV Envelope Protein gp120