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A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency.


ABSTRACT: Integral membrane proteins (IMPs) control the flow of information and nutrients across cell membranes, yet IMP mechanistic studies are hindered by difficulties in expression. We investigate this issue by addressing the connection between IMP sequence and observed expression levels. For homologs of the IMP TatC, observed expression levels vary widely and are affected by small changes in protein sequence. The effect of sequence changes on experimentally observed expression levels strongly correlates with the simulated integration efficiency obtained from coarse-grained modeling, which is directly confirmed using an in vivo assay. Furthermore, mutations that improve the simulated integration efficiency likewise increase the experimentally observed expression levels. Demonstration of these trends in both Escherichia coli and Mycobacterium smegmatis suggests that the results are general to other expression systems. This work suggests that IMP integration is a determinant for successful expression, raising the possibility of controlling IMP expression via rational design.

SUBMITTER: Marshall SS 

PROVIDER: S-EPMC5001948 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature

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A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency.

Marshall Stephen S SS   Niesen Michiel J M MJM   Müller Axel A   Tiemann Katrin K   Saladi Shyam M SM   Galimidi Rachel P RP   Zhang Bin B   Clemons William M WM   Miller Thomas F TF  

Cell reports 20160811 8


Integral membrane proteins (IMPs) control the flow of information and nutrients across cell membranes, yet IMP mechanistic studies are hindered by difficulties in expression. We investigate this issue by addressing the connection between IMP sequence and observed expression levels. For homologs of the IMP TatC, observed expression levels vary widely and are affected by small changes in protein sequence. The effect of sequence changes on experimentally observed expression levels strongly correlat  ...[more]

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