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

JOURNAL OF MOLECULAR BIOLOGY

卷:411

Article

Bonsor, Daniel A.¹  Poste, Sandra¹  Pierce, Brian G.²  Wang, Ningyan³  Zhu, Penny¹  Buonpane, Rebecca A.³  Weng, Zhiping²  Kranz, David M.³  Sundberg, Eric J.¹ 

[1] Boston Biomed Res Inst, Watertown, MA 02472 USA

[2] Univ Massachusetts, Program Bioinformat & Integrat Biol, Sch Med, Worcester, MA 01605 USA

[3] Univ Illinois, Dept Biochem, Urbana, IL 61801 USA

关键词: protein-protein interactions;    protein engineering;    yeast display;    X-ray crystallography;    computational biology;   

DOI  :  10.1016/j.jmb.2011.06.009

来源: Elsevier

【 摘 要 】

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. (C) 2011 Elsevier Ltd. All rights reserved.

【 授权许可】

Free