Production of membrane proteins without cells or detergents.
The production of membrane proteins in cellular systems is besieged by several problems due to their hydrophobic nature which often causes misfolding, protein aggregation and cytotoxicity, resulting in poor yields of stable proteins. Cell-free expression has emerged as one of the most versatile alternatives for circumventing these obstacles by producing membrane proteins directly into designed hydrophobic environments. Efficient optimisation of expression and solubilisation conditions using a variety of detergents, membrane mimetics and lipids has yielded structurally and functionally intact membrane proteins, with yields several fold above the levels possible from cell-based systems. Here we review recently developed techniques available to produce functional membrane proteins, and discuss amphipols, nanodisc and styrene maleic acid lipid particle (SMALP) technologies that can be exploited alongside cell-free expression of membrane proteins.
Rajesh
Sundaresan
S
School of Cancer Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
Knowles
Timothy
T
Overduin
Michael
M
D002474:Cell-Free System / Q000378:metabolism*; D003902:Detergents / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D017510:Protein Folding; D014908:Triticum / Q000196:embryology
D003902:Detergents; D008565:Membrane Proteins