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Folding Engineering Strategies for Efficient Membrane Protein Production in E. coli

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Therapeutic Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 899))

Abstract

Membrane proteins are notoriously difficult to produce at the high levels required for structural and biochemical characterization. Among the various expression systems used to date, the enteric bacterium Escherichia coli remains one of the best characterized and most versatile. However, membrane protein overexpression in E. coli is often accompanied by toxicity and low yields of functional product. Here, we briefly review the involvement of signal recognition particle, trigger factor, and YidC in α-helical membrane protein biogenesis and describe a set of strains, vectors, and chaperone co-expression plasmids that can lead to significant gains in the production of recombinant membrane proteins in E. coli. Methods to quantify membrane proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis are also provided.

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Acknowledgments

BLN gratefully acknowledges NSF-IGERT fellowship support from the University of Washington Center for Nanotechnology. This work was supported by NSF award BBBE-0854511.

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Authors and Affiliations

  1. Department of Chemical Engineering, University of Washington, Seattle, WA, USA

    Brent L. Nannenga & François Baneyx

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  1. Brent L. Nannenga
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  2. François Baneyx
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Correspondence to François Baneyx .

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  1. MedImmune, LLC, MedImmune Way One, Gaithersburg, 20878, USA

    Vladimir Voynov

  2. , Department of Physical Biochemistry, Biogen Idec, Cambridge Center 14, Cambridge, 02142, Massachusetts, USA

    Justin A. Caravella

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Nannenga, B.L., Baneyx, F. (2012). Folding Engineering Strategies for Efficient Membrane Protein Production in E. coli . In: Voynov, V., Caravella, J. (eds) Therapeutic Proteins. Methods in Molecular Biology, vol 899. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-921-1_12

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  • DOI: https://doi.org/10.1007/978-1-61779-921-1_12

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