This case study shows the positive impact of the right chemical compounds on functional stabilization of membrane proteins (WO2015158575). We investigated the ability of novel glycosylated amphiphilic calixarene-based detergent (Calxglycoside) to stabilize already solubilized and purified Bacteriorhodopsin (bR). The protein was obtained from commercial source and then complemented with different compounds octyl glucoside (OG), Lauryl Maltose Neopentyl Glycol (LMNG), Amphipol (A835), Facial Amphiphile 3 (FA-3) and Calxglycoside. The activity of different bR preparations was assessed by absorbance measurement at 560nm over time. We showed that significant difference on stability was observed among the conditions.
Interestingly, the reference detergent OG which was used to solubilize and purify the protein, do not allow to maintain the bR activity overtime and resulted on a basal activity of 25% after only couple of days. Amphipol (A835) was also not a very good condition. Facial amphiphile FA3 and LMNG did maintain better bR activity in comparison to OG or A835. Interestingly in comparison to other compounds, Calxglycoside did maintain the bR activity up to 60-70% after 45 days at room temperature and 35 more days at 37°C.
Thus, these results indicate that Calxglycoside is substantially better at stabilizing the bR than other conventional or more recent commercialized detergents or surfactants.
Off-the-shelf functional and pure Bacteriorhodopsin protein produced as described above is now available for sale in our catalog.
Dauvergne J. et al. 2019