A dual-reporter system for investigating and optimizing protein translation and folding in E. coli.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
19 10 2021
19 10 2021
Historique:
received:
13
01
2020
accepted:
01
10
2021
entrez:
20
10
2021
pubmed:
21
10
2021
medline:
1
12
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 34667164
doi: 10.1038/s41467-021-26337-1
pii: 10.1038/s41467-021-26337-1
pmc: PMC8526717
doi:
Substances chimiques
Escherichia coli Proteins
0
Luminescent Proteins
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6093Informations de copyright
© 2021. The Author(s).
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