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Nat Commun. 2015 Apr 8;6:6826. doi: 10.1038/ncomms7826.

Making water-soluble integral membrane proteins in vivo using an amphipathic protein fusion strategy.

Author information

1
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York 14853, USA.
2
School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.
3
Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109, USA.
4
Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14850, USA.
5
Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.

Abstract

Integral membrane proteins (IMPs) play crucial roles in all cells and represent attractive pharmacological targets. However, functional and structural studies of IMPs are hindered by their hydrophobic nature and the fact that they are generally unstable following extraction from their native membrane environment using detergents. Here we devise a general strategy for in vivo solubilization of IMPs in structurally relevant conformations without the need for detergents or mutations to the IMP itself, as an alternative to extraction and in vitro solubilization. This technique, called SIMPLEx (solubilization of IMPs with high levels of expression), allows the direct expression of soluble products in living cells by simply fusing an IMP target with truncated apolipoprotein A-I, which serves as an amphipathic proteic 'shield' that sequesters the IMP from water and promotes its solubilization.

PMID:
25851941
PMCID:
PMC4403311
DOI:
10.1038/ncomms7826
[Indexed for MEDLINE]
Free PMC Article

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