Molecular basis of a million-fold affinity maturation process in a protein-protein interaction

Daniel A. Bonsor, Sandra Postel, Brian G. Pierce, Ningyan Wang, Penny Zhu, Rebecca A. Buonpane, Zhiping Weng, David M Kranz, Eric J. Sundberg

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)321-328
Number of pages8
JournalJournal of Molecular Biology
Volume411
Issue number2
DOIs
StatePublished - Aug 12 2011

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Protein Engineering
Proteins
Superantigens
X Ray Crystallography
Firearms
T-Cell Antigen Receptor
Mutagenesis
Pharmaceutical Preparations
Therapeutics

Keywords

  • X-ray crystallography
  • computational biology
  • protein engineering
  • protein-protein interactions
  • yeast display

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Bonsor, D. A., Postel, S., Pierce, B. G., Wang, N., Zhu, P., Buonpane, R. A., ... Sundberg, E. J. (2011). Molecular basis of a million-fold affinity maturation process in a protein-protein interaction. Journal of Molecular Biology, 411(2), 321-328. https://doi.org/10.1016/j.jmb.2011.06.009

Molecular basis of a million-fold affinity maturation process in a protein-protein interaction. / Bonsor, Daniel A.; Postel, Sandra; Pierce, Brian G.; Wang, Ningyan; Zhu, Penny; Buonpane, Rebecca A.; Weng, Zhiping; Kranz, David M; Sundberg, Eric J.

In: Journal of Molecular Biology, Vol. 411, No. 2, 12.08.2011, p. 321-328.

Research output: Contribution to journalArticle

Bonsor, DA, Postel, S, Pierce, BG, Wang, N, Zhu, P, Buonpane, RA, Weng, Z, Kranz, DM & Sundberg, EJ 2011, 'Molecular basis of a million-fold affinity maturation process in a protein-protein interaction', Journal of Molecular Biology, vol. 411, no. 2, pp. 321-328. https://doi.org/10.1016/j.jmb.2011.06.009
Bonsor, Daniel A. ; Postel, Sandra ; Pierce, Brian G. ; Wang, Ningyan ; Zhu, Penny ; Buonpane, Rebecca A. ; Weng, Zhiping ; Kranz, David M ; Sundberg, Eric J. / Molecular basis of a million-fold affinity maturation process in a protein-protein interaction. In: Journal of Molecular Biology. 2011 ; Vol. 411, No. 2. pp. 321-328.
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