Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure can provide specific custom Mempro™ Bet v1-like protein production services using detergent-free expression system.
Bet v1-like superfamily proteins are a group of proteins containing a beta-alpha-beta-alpha domain, which can be known as the white birch major pollen allergen. Bet v1-like proteins are the lipocalin-like proteins that play crucial roles in lipid transport and pathogenesis as well as other biological functions, such as biosynthesis of secondary metabolites, degradation of aromatic compounds, and antibiotic resistance. Members of Bet v1-like superfamily possess the TATA binding protein (TBP)-like fold that containing beta-alpha-beta(4)-alpha structural domain. Conventional techniques for membrane protein production require extraction from the membrane into detergent micelles. However, detergent can induce conformational changes on the membrane proteins. Our substitutive detergent-free expression system can best preserve the native conformation of Bet v1-like proteins for further structural analysis.
Figure 1. A structural model of STAR-related lipid transfer protein from bacteria. (OPM database)
Creative Biostructure has extensive expertise in high-yield Bet v1-like protein production in detergent-free membrane protein expression system, we can utilize various strategies for Mempro™ detergent-free Bet v1-like protein production, including:
Nanodiscs are the self-assembled nanotechnology to stabilize transmembrane proteins removed from the membrane by membrane scaffold proteins (MSPs) in absence of detergent. Creative Biostructure can reconstitute Bet v1-like proteins into nanodiscs in detergent-free procedure: Expressing the Bet v1-like proteins in the cell-free expression system with the addition of pre-assembled nanodiscs.
The amphipathic amphipols have the ability to enwrap transmembrane proteins, allowing them to stay folded. Creative Biostructure can apply A8-35 to solubilize Bet v1-like proteins during purification and stabilization processes.
The SMALPs can be self-assembled by the simple addition of the SMA co-polymers. At neutral or alkaline pH condition, the disc-like structure assembles itself, encapsulating Bet v1-like proteins in a formation amenable to be purified.
These novel detergent-free techniques for membrane protein production can be obtained easily, and enabling more comprehensively structural and functional studies.
Creative Biostructure provides other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
C. Tribet, et al. (1996). Amphipols: Polymers that keep membrane proteins soluble in aqueous solutions. Proc Natl Acad Sci U. S. A. 93: 15047-15050.
F. Roth-Walter, et al. (2014). Bet v1 from birch pollen is a lipocalin-like protein acting as Allergen only when devoid of iron by promoting Th2 lymphocytes. J. Biol. Chem., 289: 17416-17421.
J. Chakrabortya and T. K. Duttaa (2011). From lipid transport to oxygenation of aromatic compounds: evolution within the bet v1-like superfamily. J. Biomol. Struc. Dynam., 29(1): 67-78.
M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans., 39: 813-818.