7AON

Crystal structure of CI2 double mutant L49I,I57V

Summary for 7AON

• Entry DOI: 10.2210/pdb7aon/pdb
• Related: 2CI2 7A1H 7AOK
• Descriptor: Subtilisin-chymotrypsin inhibitor-2A, SULFATE ION, GLYCEROL, ... (4 entities in total)
• Functional Keywords: protease inhibitor, protein binding
• Biological source: Hordeum vulgare (Barley)
• Total number of polymer chains: 1
• Total formula weight: 7583.82

Authors

Olsen, J.G.,Teilum, K.,Hamborg, L.,Roche, J.V. (deposition date: 2020-10-14, release date: 2020-12-09, Last modification date: 2024-01-31)

Primary citation

Hamborg, L.,Granata, D.,Olsen, J.G.,Roche, J.V.,Pedersen, L.E.,Nielsen, A.T.,Lindorff-Larsen, K.,Teilum, K.
Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli.
Commun Biol, 4:980-980, 2021

PubMed Abstract: Most single point mutations destabilize folded proteins. Mutations that stabilize a protein typically only have a small effect and multiple mutations are often needed to substantially increase the stability. Multiple point mutations may act synergistically on the stability, and it is often not straightforward to predict their combined effect from the individual contributions. Here, we have applied an efficient in-cell assay in E. coli to select variants of the barley chymotrypsin inhibitor 2 with increased stability. We find two variants that are more than 3.8 kJ mol more stable than the wild-type. In one case, the increased stability is the effect of the single substitution D55G. The other case is a double mutant, L49I/I57V, which is 5.1 kJ mol more stable than the sum of the effects of the individual mutations. In addition to demonstrating the strength of our selection system for finding stabilizing mutations, our work also demonstrate how subtle conformational effects may modulate stability.

PubMed: 34408246
DOI: 10.1038/s42003-021-02490-7

Experimental method

X-RAY DIFFRACTION (1.3 Å)