Abstract
In addition to the cytoplasmic membrane, Gram-negative bacteria have a second lipid bilayer, the outer membrane, which is the de facto barrier between the cell and the extracellular milieu. Virtually all integral proteins of the outer membrane form β-barrels, which are inserted into the outer membrane by the BAM complex. Some outer membrane proteins, like the porins and trimeric autotransporter adhesins, are multimeric. In the former case, the porin trimer consists of three individual β-barrels, whereas in the latter, the single autotransporter β-barrel domain is formed by three separate polypeptides. This chapter reviews methods to investigate the folding and membrane insertion of multimeric OMPs and further explains the use of a BamA depletion strain to study the effects of the BAM complex on multimeric OMPs in E. coli.
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Acknowledgements
We are grateful to Prof. Tracy Palmer, University of Dundee, for providing us with the BamA depletion strain. For funding, we thank FEMS (an advanced fellowship to J.C.L.), UKT fortüne program (F1433253 to P.O.) and the German Science Foundation (SFB766/B10 to D.L.). We wish to thank Prof. Andrei Lupas (Max Planck Institute for Developmental Biology, Tübingen), and Dr. Monika Schütz and Prof. Ingo B. Autenrieth (University Clinics Tübingen) for continuing support and collaboration.
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Leo, J.C., Oberhettinger, P., Linke, D. (2015). Assessing the Outer Membrane Insertion and Folding of Multimeric Transmembrane β-Barrel Proteins. In: Buchanan, S., Noinaj, N. (eds) The BAM Complex. Methods in Molecular Biology, vol 1329. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2871-2_12
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DOI: https://doi.org/10.1007/978-1-4939-2871-2_12
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