Mempro™ Cell-Free Phosphatidylinositol 3-/4-Kinase Production

Creative Biostructure provides costum Mempro™ protein production services for phosphatidylinositol 3-/4-kinase (PI 3-/4-kinase) based on cell-free expression system.

Mempro™ cell-free protein production system is the innovated system to produce membrane proteins, which can bypass the limitations that encountered in cell-based production system, such as host cell toxicity, low yield and solubilization and purification using detergents. PI 3-/4-kinases are a family of enzymes that play crucial roles in cellular functions such as cell growth, proliferation, differentiation, motility, survival and intracellular trafficking, which in turn are involved in cancer. PI3-kinases are the related intracellular signal transducer enzyme that can phosphorylate phosphoinositides on the 3-hydroxyl group of the inositol ring and function as secondary messengers in cell signaling. PI4-kinases are the enzymes that act on PI in the first committed step in the production of the secondary messenger inositol-1'4'5'-trisphosphate. Notably, PI 3-/4-kinases share a conserved catalytic domain at their C-terminal section.

Figure 1. PIK-93 inhibitor (yellow) interacts with the PI 3-Kinase 110 gamma subunit. (Wikipedia)

Creative Biostructure has extensive expertise in high-yield PI 3-/4-kinase production using cell-free protein expression system, we can perform 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 membrane protein production systems, E. coli S30 extract is widely used for PI 3-/4-kinase production. Wheat germ extract can provide eukaryotic translation system for PI 3-/4-kinase expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for PI 3-/4-kinase expression. Rabbit reticulocyte lysate is an important tool for eukaryotic cell-free membrane protein expression. To employ an appropriate source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.

With the Mempro™ cell-free protein production platform, Creative Biostructure are capable of expressing, purifying and crystallizing PI 3-/4-kinase to perform further structural and functional studies.

Creative Biostructure provides other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
Cell-free protein synthesis. (https://en.wikipedia.org/wiki/Cell-free_protein_synthesis)
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
J. Y. Jung, et al. (2002). Phosphatidylinositol 3- and 4-phosphate are required for normal stomatal movements. Plant Cell., 14(10): 2399-2412.
M. Fujimoto, et al. (2015). Phosphatidylinositol 3-kinase and 4-kinase have distinct roles in intracellular trafficking of cellulose synthase complexes in Arabidopsis thaliana. Plant Cell Physiol., 56(2): 287-298.