A dual-reporter system for investigating and optimizing protein translation and folding in E. coli
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A dual-reporter system for investigating and optimizing protein translation and folding in E. coli. / Zutz, Ariane; Hamborg, Louise; Pedersen, Lasse Ebdrup; Kassem, Maher M.; Papaleo, Elena; Koza, Anna; Herrgård, Markus J.; Ingemann Jensen, Sheila; Teilum, Kaare; Lindorff-Larsen, Kresten; Nielsen, Alex Toftgaard.
I: Nature Communications, Bind 12, 6093, 2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - A dual-reporter system for investigating and optimizing protein translation and folding in E. coli
AU - Zutz, Ariane
AU - Hamborg, Louise
AU - Pedersen, Lasse Ebdrup
AU - Kassem, Maher M.
AU - Papaleo, Elena
AU - Koza, Anna
AU - Herrgård, Markus J.
AU - Ingemann Jensen, Sheila
AU - Teilum, Kaare
AU - Lindorff-Larsen, Kresten
AU - Nielsen, Alex Toftgaard
N1 - Publisher Copyright: © 2021, The Author(s).
PY - 2021
Y1 - 2021
N2 - Strategies for investigating and optimizing the expression and folding of proteins for biotechnological and pharmaceutical purposes are in high demand. Here, we describe a dual-reporter biosensor system that simultaneously assesses in vivo protein translation and protein folding, thereby enabling rapid screening of mutant libraries. We have validated the dual-reporter system on five different proteins and find an excellent correlation between reporter signals and the levels of protein expression and solubility of the proteins. We further demonstrate the applicability of the dual-reporter system as a screening assay for deep mutational scanning experiments. The system enables high throughput selection of protein variants with high expression levels and altered protein stability. Next generation sequencing analysis of the resulting libraries of protein variants show a good correlation between computationally predicted and experimentally determined protein stabilities. We furthermore show that the mutational experimental data obtained using this system may be useful for protein structure calculations.
AB - Strategies for investigating and optimizing the expression and folding of proteins for biotechnological and pharmaceutical purposes are in high demand. Here, we describe a dual-reporter biosensor system that simultaneously assesses in vivo protein translation and protein folding, thereby enabling rapid screening of mutant libraries. We have validated the dual-reporter system on five different proteins and find an excellent correlation between reporter signals and the levels of protein expression and solubility of the proteins. We further demonstrate the applicability of the dual-reporter system as a screening assay for deep mutational scanning experiments. The system enables high throughput selection of protein variants with high expression levels and altered protein stability. Next generation sequencing analysis of the resulting libraries of protein variants show a good correlation between computationally predicted and experimentally determined protein stabilities. We furthermore show that the mutational experimental data obtained using this system may be useful for protein structure calculations.
U2 - 10.1038/s41467-021-26337-1
DO - 10.1038/s41467-021-26337-1
M3 - Journal article
C2 - 34667164
AN - SCOPUS:85113144928
VL - 12
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
M1 - 6093
ER -
ID: 283214592