Folding engineering strategies for efficient membrane protein production in E. coli

Brent L. Nannenga, François Baneyx

Research output: Chapter in Book/Report/Conference proceedingChapter

6 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationTherapeutic Proteins
Subtitle of host publicationMethods and Protocols
EditorsVoynov Vladimir, Caravella Justin
Pages187-202
Number of pages16
DOIs
StatePublished - Jul 27 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume899
ISSN (Print)1064-3745

Keywords

  • Insertase
  • Molecular chaperone
  • Signal recognition particle
  • Trigger factor
  • YidC

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Nannenga, B. L., & Baneyx, F. (2012). Folding engineering strategies for efficient membrane protein production in E. coli. In V. Vladimir, & C. Justin (Eds.), Therapeutic Proteins: Methods and Protocols (pp. 187-202). (Methods in Molecular Biology; Vol. 899). https://doi.org/10.1007/978-1-61779-921-1_12