Based on the comprehensive membrane protein production platform established through years of experience, scientists form Creative Biostructure can offer tailored Mempro™ PLC-like phosphodiesterase production services using detergent-free expression system.
Phosphodiesterases are known as cyclic nucleotide phosphodiesterases, which have important clinical roles. Phospholipase C (PLC) enzymes usually play an important role in kinds of signalling cascades. Various of the crucial biology processes were regulated by cyclic nucleotide phosphodiesterases, such as duration, localization and amplitude of cyclic nucleotide signaling. cAMP and cGMP which are second messenger molecules degraded by a flock of enzymes were made up of cyclic nucleotide phosphodiesterases.
PLC isozymes are responsible to the hydrolyzation of phosphatidylinositol 4,5-bisphosphate and divided into second messengers diacylglycerol and inositol 1,4,5-trisphosphate. The reaction is directly activated by Ras-like GTPases and heterotrimeric G proteins. There are plenty of PLC-like phosphodiesterase, including bacterial phosphatidylinositol-specific phospholipase C, glycerophosphodiester phosphodiesterases and mammalian phospholipase C isozyme D1.
Figure: Cell-free synthesis of membrane proteins in presence of different lipid based membrane structures.
Creative Biostructure has exteinsive expertise in high-quality PLC-like phosphodiesterase production using detergent-free membrane protein expression system, we can employ various strategies for Mempro™ detergent-free protein production, including:
Nanodiscs are soluble nanoscale phospholipid bilayers, which are composed of membrane scaffold proteins (MSPs). Integral membrane proteins are able to assemble themselves by nanodiscs which are general used for structural, enzymatic or biophysical researches. Nanodiscs enable to maintain membrane proteins in solution, and offer an inartificial nature phospholipid bilayer environment in which the incorporated target keeps stably.
Amphipols are absolutely serviceable tools for handling membrane proteins in detergent-free aqueous solutions, and essentially they are a new class of surfactants. Amphipols can be characteristically used in biophysics and biochemistry by the physicochemical properties of amphipols cut, which is based on the wide resources of polymer chemistry.
SMALPs are disc-like structures containing a protein in a lipid bilayer, the use of styrene maleic acid copolymer (SMA) can make the membrane proteins and retention segregate in the inartificial lipid environment. Compared with nanodiscs, bicelles and amphipols, the most significantly advantage of SMALPs is that there is no need to extraction of proteins from membranes with possibly destabilising detergents.
These novel detergent-free approaches for PLC-like phosphodiesterase production can be obtained easily, enabling more comprehensively 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:R. K. Verma, et al. (2015). Major intrinsic protein superfamily: channels with unique structural features and diverse selectivity filters. Methods Enzymol. 557: 485-520.E. Meldrum, et al. (1991)., The PtdIns-PLC superfamily and signal transduction. Biochim. 54:49-71. Jump up, et al. (1992)., Multiple forms of phospholipase C isozymes and their activation mechanisms.Second Messenger Phosphoprotein. 26: 35-61 M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans.39: 813-818. N. Yan (2015). Structural biology of the major facilitator superfamily transporters. Biophys., 44: 257-283.