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Molecular Basis of a Million-Fold Affinity Maturation Process in a Protein-Protein Interaction

Journal Article · · Journal of Molecular Biology
Protein engineering is becoming increasingly important for pharmaceutical applications where controlling the specificity and affinity of engineered proteins is required to create targeted protein therapeutics. Affinity increases of several thousand-fold are now routine for a variety of protein engineering approaches, and the structural and energetic bases of affinity maturation have been investigated in a number of such cases. Previously, a 3-million-fold affinity maturation process was achieved in a protein-protein interaction composed of a variant T-cell receptor fragment and a bacterial superantigen. Here, we present the molecular basis of this affinity increase. Using X-ray crystallography, shotgun reversion/replacement scanning mutagenesis, and computational analysis, we describe, in molecular detail, a process by which extrainterfacial regions of a protein complex can be rationally manipulated to significantly improve protein engineering outcomes.
Research Organization:
BROOKHAVEN NATIONAL LABORATORY (BNL)
Sponsoring Organization:
USDOE SC OFFICE OF SCIENCE (SC)
DOE Contract Number:
AC02-98CH10886
OSTI ID:
1041876
Report Number(s):
BNL--97554-2012-JA
Journal Information:
Journal of Molecular Biology, Journal Name: Journal of Molecular Biology Journal Issue: 2 Vol. 411; ISSN JMOBAK; ISSN 0022-2836
Country of Publication:
United States
Language:
English