Improving membrane protein expression and function using genomic edits

Expression of membrane proteins often leads to growth inhibition and perturbs central metabolism and this burden varies with the protein being overexpressed. There are also known strain backgrounds that allow greater expression of membrane proteins but that differ in efficacy across proteins. We hyp...

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Bibliographic Details
Published in:Scientific reports 2017-10, Vol.7 (1), p.13030-14, Article 13030
Main Authors: Jensen, Heather M., Eng, Thomas, Chubukov, Victor, Herbert, Robin A., Mukhopadhyay, Aindrila
Format: Article
Language:English
Subjects:
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Applied microbiology
BASIC BIOLOGICAL SCIENCES
E coli
Genome editing
Genomes
Humanities and Social Sciences
Insertion
Membrane proteins
Membranes
Molecular biology
multidisciplinary
Protein expression
Proteins
Science
Science (multidisciplinary)
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Summary:Expression of membrane proteins often leads to growth inhibition and perturbs central metabolism and this burden varies with the protein being overexpressed. There are also known strain backgrounds that allow greater expression of membrane proteins but that differ in efficacy across proteins. We hypothesized that for any membrane protein, it may be possible to identify a modified strain background where its expression can be accommodated with less burden. To directly test this hypothesis, we used a bar-coded transposon insertion library in tandem with cell sorting to assess genome-wide impact of gene deletions on membrane protein expression. The expression of five membrane proteins (CyoB, CydB, MdlB, YidC, and LepI) and one soluble protein (GST), each fused to GFP, was examined. We identified Escherichia coli mutants that demonstrated increased membrane protein expression relative to that in wild type. For two of the proteins (CyoB and CydB), we conducted functional assays to confirm that the increase in protein expression also led to phenotypic improvement in function. This study represents a systematic approach to broadly identify genetic loci that can be used to improve membrane protein expression, and our method can be used to improve expression of any protein that poses a cellular burden.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-017-12901-7