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

Brent Nannenga, François Baneyx

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Citations (Scopus)

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 publicationMethods in Molecular Biology
Pages187-202
Number of pages16
Volume899
DOIs
StatePublished - 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume899
ISSN (Print)10643745

Fingerprint

Membrane Proteins
Escherichia coli
Signal Recognition Particle
Enterobacteriaceae
Recombinant Proteins
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Plasmids

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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

Folding engineering strategies for efficient membrane protein production in E. coli. / Nannenga, Brent; Baneyx, François.

Methods in Molecular Biology. Vol. 899 2012. p. 187-202 (Methods in Molecular Biology; Vol. 899).

Research output: Chapter in Book/Report/Conference proceedingChapter

Nannenga, B & Baneyx, F 2012, Folding engineering strategies for efficient membrane protein production in E. coli. in Methods in Molecular Biology. vol. 899, Methods in Molecular Biology, vol. 899, pp. 187-202. https://doi.org/10.1007/978-1-61779-921-1_12
Nannenga B, Baneyx F. Folding engineering strategies for efficient membrane protein production in E. coli. In Methods in Molecular Biology. Vol. 899. 2012. p. 187-202. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-921-1_12
Nannenga, Brent ; Baneyx, François. / Folding engineering strategies for efficient membrane protein production in E. coli. Methods in Molecular Biology. Vol. 899 2012. pp. 187-202 (Methods in Molecular Biology).
@inbook{21b91ad317964ee59ab596d01f289420,
title = "Folding engineering strategies for efficient membrane protein production in E. coli",
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.",
keywords = "Insertase, Molecular chaperone, Signal recognition particle, Trigger factor, YidC",
author = "Brent Nannenga and Fran{\cc}ois Baneyx",
year = "2012",
doi = "10.1007/978-1-61779-921-1_12",
language = "English (US)",
isbn = "9781617799204",
volume = "899",
series = "Methods in Molecular Biology",
pages = "187--202",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Folding engineering strategies for efficient membrane protein production in E. coli

AU - Nannenga, Brent

AU - Baneyx, François

PY - 2012

Y1 - 2012

N2 - 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.

AB - 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.

KW - Insertase

KW - Molecular chaperone

KW - Signal recognition particle

KW - Trigger factor

KW - YidC

UR - http://www.scopus.com/inward/record.url?scp=84864127248&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864127248&partnerID=8YFLogxK

U2 - 10.1007/978-1-61779-921-1_12

DO - 10.1007/978-1-61779-921-1_12

M3 - Chapter

C2 - 22735954

AN - SCOPUS:84864127248

SN - 9781617799204

VL - 899

T3 - Methods in Molecular Biology

SP - 187

EP - 202

BT - Methods in Molecular Biology

ER -