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N Biotechnol. 2011 Apr 30;28(3):250-4. doi: 10.1016/j.nbt.2010.07.011. Epub 2010 Jul 21.

Production of membrane proteins without cells or detergents.

Author information

1
School of Cancer Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.

Abstract

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.

PMID:
20654746
DOI:
10.1016/j.nbt.2010.07.011
[Indexed for MEDLINE]

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