Creative Biostructure has established a comprehensive membrane protein production platform based on years of experience, our scientists can provide custom Mempro™ transmembrane protein PLC-like phosphodiesterase production services by adopting cell-based expression system.
Mempro™ cell-based protein production system is the one of our most applied and advanced method for membrane proteins production. Phosphoinositide phospholipase C (PLC) is a eukaryotic intracellular membrane protein family of enzymes. PLCs are essential for signal transduction processes through calcium-protein kinase C signaling pathway regulation, and lead to various bioprocess including cell proliferation, differentiation, adhesion, and secretion. There are 12 PLC isoforms have been cloned, and are classified into five groups considering the sequence homology and the activation mechanism up to date. Activation of PLC-like phosphodiesterases is directly induced by heterotrimeric G proteins and Ras-like GTPases. PLC-like phosphodiesterases can induce the phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolyzation into the second messengers diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3).
Figure 1. Structural Details of the Active Site of PLC-β2. (Molecular Cell, 2008)
Creative Biostructure can offer high-quality membrane protein production services, and we can employ a lot of Mempro™ cell-based protein production systems including:
• Mempro™ PLC-like Phosphodiesterase Production using Bacterial Cells SystemEscherichia coli (E. coli) is the most widely applied bacterial host for membrane protein production. Lemo21 (DE3) strain is optimized for our Mempro™ protein production in our bacterial cells system. Whole-cell fluorescence can be used to monitor the membrane protein-GFP fusions production by Lemo21 (DE3), which enables the integrity of the production. The fluorescence assay also facilitates the choice of an optimally detergent to solubilize the target from the membrane, and to monitor the stability of this target membrane proteins within this detergent.
• Mempro™ PLC-like Phosphodiesterase Production using Insect Cells SystemThe most applied system for eukaryotic membrane protein production is in cultured insect cells and larvae. Mempro™ protein production using insect cells system can increase the expression level and reduce the truncated proteins compared to E. coli system according to similar codon usage rules. Our innovational insect cells system based on baculovirus is advanced in several aspects as below：Accuracy;Easy scale-up;High membrane protein yield;Proper folding and post translation modifications;Use of cell lines ideal for suspension culture.
• Mempro™ PLC-like Phosphodiesterase Production using Yeast Cells SystemSingle-celled yeast is an easy, quick and economic culture host and able to apply multiple post translation modifications for eukaryotic membrane protein. The most widely used two species of yeast for membrane protein production are Pichia pastoris (P. pastoris) and Saccharomyces cerevisiae (S. cerevisiae). Creative Biostructure has developed several strategies to improve yields per cell through optimizing the expression plasmid, host cell and culture conditions.
• Mempro™ PLC-like Phosphodiesterase Production using Mammalian Cells SystemCreative Biostructure can offer another great service-Mempro™ protein production in mammalian cells system. This system helps to correct folding and post-translational modifications for membrane protein. We have proprietary Mempro™ protein production in mammalian cells system with the best structural and functional features of target eukaryotic membrane proteins from numerous mammalian cell types including human embryonic kidney cells (HEK-293), Chinese hamster ovary cells (CHO), monkey kidney fibroblast cells (COS-7) and baby hamster kidney cells (BHK-21).
Creative Biostructure provides Mempro™ cell-based protein production services from expressing, isolating, purifying to crystallizing the membrane protein PLC-like phosphodiesterase.
We provide an array of Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
References:Y. Sun, et. al. (2010). Kinetic Scaffolding Mediated by a Phospholipase C–β and GqSignaling Complex. Science, 30, 974-980.S. N. Hicks, et al. (2008). General and Versatile Autoinhibition of PLC Isozymes. Molecular. Cell, 31: 383–394. M. Nomikos (2007). Binding of phosphoinositide-specific phospholipase C-zeta (PLC-zeta) to phospholipid membranes: potential role of an unstructured cluster of basic residues. J. Biol. Chem., 282, 16644–16653.