Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli

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Standard

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.

I: Communications Biology , Bind 4, 980, 2021.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Hamborg, L, Granata, D, Olsen, JG, Roche, JV, Pedersen, LE, Nielsen, AT, Lindorff-Larsen, K & Teilum, K 2021, 'Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli', Communications Biology , bind 4, 980. https://doi.org/10.1038/s42003-021-02490-7

APA

Hamborg, L., Granata, D., Olsen, J. G., Roche, J. V., Pedersen, L. E., Nielsen, A. T., Lindorff-Larsen, K., & Teilum, K. (2021). Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli. Communications Biology , 4, [980]. https://doi.org/10.1038/s42003-021-02490-7

Vancouver

Hamborg L, Granata D, Olsen JG, Roche JV, Pedersen LE, Nielsen AT o.a. Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli. Communications Biology . 2021;4. 980. https://doi.org/10.1038/s42003-021-02490-7

Author

Hamborg, Louise ; Granata, Daniele ; Olsen, Johan G. ; Roche, Jennifer Virginia ; Pedersen, Lasse Ebdrup ; Nielsen, Alex Toftgaard ; Lindorff-Larsen, Kresten ; Teilum, Kaare. / Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli. I: Communications Biology . 2021 ; Bind 4.

Bibtex

@article{a304e20404cc485ca0e346b874ab6613,

title = "Synergistic stabilization of a double mutant in chymotrypsin inhibitor 2 from a library screen in E. coli",

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−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.",

author = "Louise Hamborg and Daniele Granata and Olsen, {Johan G.} and Roche, {Jennifer Virginia} and Pedersen, {Lasse Ebdrup} and Nielsen, {Alex Toftgaard} and Kresten Lindorff-Larsen and Kaare Teilum",

note = "Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

doi = "10.1038/s42003-021-02490-7",

language = "English",

volume = "4",

journal = "Communications Biology",

issn = "2399-3642",

publisher = "nature publishing group",

}

RIS

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

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

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