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Mempro™ PLC-like Phosphodiesterase Production Using Virus-Like Particles

Creative Biostructure now can provide custom Mempro™ PLC-like phosphodiesterase production services using virus-like particles with years of experience in membrane protein production.

Phospholipase C-like phosphodiesterase (PLC-like phosphodiesterase) is a protein superfamily contains 3 protein families: the mammalian PLC family, the bacteria PLC family and the glycerophosphoryl diester phosphodiesterase family. The three families contain various PLCs and their isozymes. PLC catalyzes the hydrolysis of phosphatidylinositol 4, 5-bisphosphate (PIP2) selectively on the glycerol side of the phosphodiester bond. Tens of cellular process are regulated by this reaction directly and hundreds more are infected indirectly. The PLC catalyzed reaction is the major pass way for PIP2, which acts as an allosteric regulator or membrane anchor for various membrane proteins. What's more, PLC is also the creator of inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG), which are important second messengers with further regulatory and metabolic functions.

Mempro™ PLC-like Phosphodiesterase Production Using Virus-Like Particles Figure 1. The schematic structure of human phospholipase C-beta isoform 2. (OPM Database)

Virus-like particles (VLPs) are high-organized, self-assembled subunits described as "empty viral protein shells" of virus. VLPs are assembled by viral structural proteins and can mimic conformation and organization of the native virus. However, VLPs are non-infectious as all of the VLPs lack of virus genomes. VLPs have been used widely in vaccine developments, antigen deliveries and recombinant membrane protein productions. Multiple virus families have been reported to be used for VLPs production, including parvoviridae, bacteriophages, retroviridae, etc.

Creative Biostructure has developed multiple host systems for expression of membrane proteins and production of VLPs to meet different experimental requirements. These host systems including but not limited to:

Green plants (tobacco, alfalfa, potato, etc.)
Bacteria (E. coli)
Insect cells
Mammalian cells

Creative Biostructure now can provide the stable PLC-like phosphodiesterases with high yield and high purity. Our scientists will co-express the target and the virus structural core polyproteins to obtain PLC-like phosphodiesterases specific lipoparticles, which are VLPs containing high concentration of specific PLC-like phosphodiesterases within their construction. Viral core proteins perform self-assembling at the plasma, and bud-off from the plasma membrane as lipoparticles. Expressed recombinant PLC-like phosphodiesterases will be captured inside the lipoparticles. We shall harvest the lipoparticles from the surfaces of host cells. Through this way, PLC-like phosphodiesterases can be isolated without detergents or mechanical disruption, so the native structure and orientation of membrane proteins can be retained within the lipoparticles.

Creative Biostructure also can provide other native or recombinant membrane proteins production service using various advanced techniques. Welcome to contact us for more details.

Ganesh Kadamur, et al. (2013).  Mammalian Phospholipase C. Annu. Rev. Physiol. 2013. 75:127–54
A.Roldão, et al. (2010). Virus-like particles in vaccine development.Expert Rev. Vaccines, 9(10): 1149-1176.
S. Willis, et al. (2008). Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry, 47(27): 6988-6890.

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