Projects per year
The fundamental importance of membrane proteins in drug discovery has meant that membrane mimetic systems for studying membrane proteins are of increasing interest. One such system has been the amphipathic, negatively charged poly(styrene-co-maleic acid) (SMA) polymer to form "SMA Lipid Particles" (SMALPs) which have been widely adopted to solubilize membrane proteins directly from the cell membrane. However, SMALPs are only soluble under basic conditions and precipitate in the presence of divalent cations required for many downstream applications. Here, we show that the positively charged poly(styrene-co-maleimide) (SMI) forms similar nanoparticles with comparable efficiency to SMA, whilst remaining functional at acidic pH and compatible with high concentrations of divalent cations. We have performed a detailed characterization of the performance of SMI that enables a direct comparison with similar data published for SMA. We also demonstrate that SMI is capable of extracting proteins directly from the cell membrane and can solubilize functional human G-protein coupled receptors (GPCRs) expressed in cultured HEK 293T cells. "SMILPs" thus provide an alternative membrane solubilization method that successfully overcomes some of the limitations of the SMALP method.
|Number of pages||11|
|Early online date||24 May 2018|
|Publication status||Published - 14 Jun 2018|
- HEK293 Cells
- Lipid Bilayers/chemistry
- Receptors, G-Protein-Coupled/metabolism
ASJC Scopus subject areas
- Materials Science(all)
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- 1 Finished
Detergent-free extraction and purification of membrane proteins to enable structural and functional studies.
Biotechnology and Biological Sciences Research Council
1/05/13 → 30/04/16
Project: Research council