Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli
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Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli. / Hamborg, Louise; Granata, Daniele; Olsen, Johan G.; Roche, Jennifer Virginia; Pedersen, Lasse Ebdrup; Nielsen, Alex Toftgaard; Lindorff-Larsen, Kresten; Teilum, Kaare.
In: Communications Biology , Vol. 4, 980, 2021.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli
AU - Hamborg, Louise
AU - Granata, Daniele
AU - Olsen, Johan G.
AU - Roche, Jennifer Virginia
AU - Pedersen, Lasse Ebdrup
AU - Nielsen, Alex Toftgaard
AU - Lindorff-Larsen, Kresten
AU - Teilum, Kaare
N1 - Publisher Copyright: © 2021, The Author(s).
PY - 2021
Y1 - 2021
N2 - 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−1 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−1 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.
AB - 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−1 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−1 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.
U2 - 10.1038/s42003-021-02490-7
DO - 10.1038/s42003-021-02490-7
M3 - Journal article
C2 - 34408246
AN - SCOPUS:85113191253
VL - 4
JO - Communications Biology
JF - Communications Biology
SN - 2399-3642
M1 - 980
ER -
ID: 279118153