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Structure. 2017 Dec 5;25(12):1867-1874.e3. doi: 10.1016/j.str.2017.11.002.

Promiscuous Protein Binding as a Function of Protein Stability.

Author information

1
Department of Biomolecular Sciences, Weizmann Institute of Science, 76100 Rehovot, Israel.
2
Structural Proteomic Unit, Weizmann Institute of Science, 76100 Rehovot, Israel.
3
Department of Biomolecular Sciences, Weizmann Institute of Science, 76100 Rehovot, Israel. Electronic address: gideon.schreiber@weizmann.ac.il.

Abstract

Proteins have evolved to balance efficient binding of desired partners with rejection of unwanted interactions. To investigate the evolution of protein-protein interactions, we selected a random library of pre-stabilized TEM1 β-lactamase against wild-type TEM1 using yeast surface display. Three mutations were sufficient to achieve micromolar affinity binding between the two. The X-ray structure emphasized that the main contribution of the selected mutations was to modify the protein fold, specifically removing the N'-terminal helix, which consequently allowed protein coupling via a β-sheet-mediated interaction resembling amyloid interaction mode. The only selected mutation located at the interaction interface (E58V) is reminiscent of the single mutation commonly causing sickle-cell anemia. Interestingly, the evolved mutations cannot be inserted into the wild-type protein due to reduced thermal stability of the resulting mutant protein. These results reveal a simple mechanism by which undesirable binding is purged by loss of thermal stability.

KEYWORDS:

TEM1; de novo protein interaction; dimerization; evolution; mutations; promiscuity; protein-protein; specificity; thermal stability; β-sheet augmentation

PMID:
29211984
DOI:
10.1016/j.str.2017.11.002
[Indexed for MEDLINE]

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