Folding engineering strategies for efficient membrane protein production in E. coli.
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.
Department of Chemical Engineering, University of Washington, Seattle, WA, USA.
Brent L
BL
Nannenga
François
F
Baneyx
D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D026901:Membrane Transport Proteins / Q000378:metabolism; D018832:Molecular Chaperones; D019696:Peptidylprolyl Isomerase / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D018271:Signal Recognition Particle / Q000737:chemistry / Q000378:metabolism
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D018271:Signal Recognition Particle; C412443:YIDC protein, E coli; C403324:trigger factor, E coli; D019696:Peptidylprolyl Isomerase