Mempro™ Cell-Free C2 Domain-containing Protein Production

Creative Biostructure can provide customized Mempro™ protein production services for C2 domain-containing proteins using cell-free expression system.

Mempro™ cell-free protein production system is the innovative system to produce intergral membrane proteins which can bypass the limitations that encountered in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification assisted with detergents. Proteins containing C2 domain can target themselves to the cell membranes. The C2 domain is a actual Ca2+-binding motif, which is orignally found as the conserved domains in the α, β and γ isoforms of mammalian Ca2+-dependent protein kinase C (PKC). Most notably, C2 domain shows the property of binding a wide range of lipids, such as phospholipids and inositol polyphosphates.

Figure 1. A model of the C2 domain interaction with the membrane (Front. Pharmacol., 2015)

Creative Biostructure has a wealth of experience in high-yield C2 domain-containing protein production using cell-free protein expression system, we can provide various strategies for Mempro™ cell-free protein production, including:

• Mempro™ Cell-Free Protein Production in E. coli;
• Mempro™ Cell-Free Protein Production in Wheat Germ;
• Mempro™ Cell-Free Protein Production in Rabbit Reticulocyte;
• Mempro™ Cell-Free Protein Production in Baculovirus.

Among Mempro™ cell-free protein production systems, E. coli S30 extract is widely used for C2 domain-containing protein production. Wheat germ extract can provide eukaryotic translation system for the production of C2 domain-containing proteins. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for C2 domain-containing protein expression. Crude rabbit reticulocyte lysate is the most important tool used for eukaryotic cell-free membrane protein expression system.

With the Mempro™ cell-free protein production platform, Creative Biostructure is capable of expressing, purifying, solubilizing and crystallizing C2 domain-containing proteins to study interactions with other protein domains as well as protein structure. Mempro™ cell-free protein production has remarkable advantages, including：

• Very high yield;
• Low cost;
• Lack of toxicity;
• Incorporation of non-natural amino acids into protein structure;
• Relatively tolerant of additives.

Creative Biostructure can provide various custom Mempro™ membrane protein production services. Please feel free to contact us for more information.

References:
C2 domain. (https://en.wikipedia.org/wiki/C2_domain#cite_note-pmid11090628-1)
E. A. Nalefski and J.J. Falke (1996). The C2 domain calcium-binding motif: structural and functional diversity. Protein Sci., 5(12): 2375-2390.
P. M. Castle, et al. (2015). Voltage-sensing phosphatase modulation by a C2 domain. Front. Pharmacol., 6: 63.
R. Sachse, et al. (2014). Membrane protein synthesis in cell-free systems: from bio-mimetic systems to bio-membranes. FEBS Lett., 588(17): 2774-2781.