ArticleScientific reports2019
Functionality of membrane proteins overexpressed and purified from E. coli is highly dependent upon the strain.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed, 57 citations in OpenAlex.
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- Proximity Labeling of Cell Surface Proteins via Cell Surface Remodeling.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Infrared Spectroscopy of SARS-CoV-2 Viral Protein: from Receptor Binding Domain to Spike Protein.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Engineered Chimera Protein Constructs to Facilitate the Production of Heterologous Transmembrane Proteins inInternational journal of molecular sciences · 2024Review
- Daptomycin avoids drug resistance mediated by the BceAB transporter inMicrobiology spectrum · 2024Article
- Escherichia coli strains with precise domain deletions in the ribonuclease RNase E can achieve greatly enhanced levels of membrane protein production.Protein science : a publication of the Protein Society · 2024Article
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- Bacterial Envelope Fractionation.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Genetically stable kill-switch using "demon and angel" expression construct of essential genes.Frontiers in bioengineering and biotechnology · 2024Article
- The transport activity of the multidrug ABC transporter BmrA does not require a wide separation of the nucleotide-binding domains.The Journal of biological chemistry · 2024Article
- Trends in the two-component system's role in the synthesis of antibiotics by Streptomyces.Applied microbiology and biotechnology · 2023Review
- Engineered nickel bioaccumulation inEngineering in life sciences · 2023Article
- Cell Surface Engineering Enables Surfaceome Profiling.ACS chemical biology · 2023Article
- eGFP as an All-in-One Tag for Purification of Membrane Proteins.Methods in molecular biology (Clifton, N.J.) · 2023Article
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
Abstract
Overexpression of correctly folded membrane proteins is a fundamental prerequisite for functional and structural studies. One of the most commonly used expression systems for the production of membrane proteins is Escherichia coli. While misfolded proteins typically aggregate and form inclusions bodies, membrane proteins that are addressed to the membrane and extractable by detergents are generally assumed to be properly folded. Accordingly, GFP fusion strategy is often used as a fluorescent proxy to monitor their expression and folding quality. Here we investigated the functionality of two different multidrug ABC transporters, the homodimer BmrA from Bacillus subtilis and the heterodimer PatA/PatB from Streptococcus pneumoniae, when produced in several E. coli strains with T7 expression system. Strikingly, while strong expression in the membrane of several strains could be achieved, we observed drastic differences in the functionality of these proteins. Moreover, we observed a general trend in which mild detergents mainly extract the population of active transporters, whereas a harsher detergent like Fos-choline 12 could solubilize transporters irrespective of their functionality. Our results suggest that the amount of T7 RNA polymerase transcripts may indirectly but notably impact the structure and activity of overexpressed membrane proteins, and advise caution when using GFP fusion strategy.
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