Mempro™ Cell-Free PLC-like Phosphodiesterase Production

Creative Biostructure offers high-quality custom Mempro™ protein production services for PLC-like phosphodiesterasese using cell-free expression system.

Mempro™ cell-free protein production system is a robust and convenient system developed by Creative Biostructure for efficient production of membrane proteins. It holds many advantages over the traditional cell-based protein production system, including less concern of toxicity, high yield, ready-to-use, post-translational modifications, etc.

Phospholipase C (PLC) catalyzes the hydrolysis of phosphatidylinositol 4,5-bisphosphate to inositol 1,4,5-trisphosphate and diacylglycerol in inositophospholipid signaling. Base on the structure, PLC can be divided into three types: β, γ and δ. PLC-like proteins is a structural domain consisting of a TIM beta/alpha-barrel which is very similar to PLC. It has a molecular weight of 130 kDa, which is less than β and γ type while greater than δ type PLC. Like δ type PLC, PLC-L also have a PH domain which is responsible for binding with PI(4,5)P2 and Ins(1,4,5)P3. PLC-L anchors to the plasma membranes until the extracellular signal stimulates the detachment of the protein from the plasma membranes when the Ins(1,4,5)P3 increases. In addition, PLC-L phosphodiesterases also contains X and Y domain and C2 region as they are in δ type PLC.

Figure 1. Crystal structures of glycerophosphodiester phosphodiesterase.

Creative Biostructure is very experienced in high-yield PLC-like phosphodiesterases production using cell-free protein expression systems, various strategies are available 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 system is the optimal system for PLC-like phosphodiesterase production. It has been successfully expressed in E. coli system. E. coli S30 extract system contains the translational machinery required for both prokaryotic and eukaryotic protein. It’s widely applied to a variety of membrane proteins for its fast and active production. Furthermore, using E. coli extract has less possibility of cross-reactivity or interference with the endogenous proteins in eukaryotic system.

Creative Biostructure is a premiere source of cell free protein expression. With the Mempro™ cell-free protein production platform, purified PLC-like phosphodiesterases could be produced in a cost-effective manner, which will ensure the progress of your research.  

We also provide other types of Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
Otsuki M, Fukami K, Kohno T, et al. Identification and characterization of a new phospholipase C-like protein, PLC-L 2[J]. Biochemical and biophysical research communications, 1999, 266(1): 97-103.