Creative Biostructure can provide excellent custom Mempro™ cytochrome P450 (CYP) production services using detergent-free expression system. We are capable of preserving the native conformation of CYPs for your structural and functional studies.
CYPs, a superfamily of heme proteins containing a heme-iron center, which are abundant in liver. CYPs play a critical role in repair of organ injury by various insults. CYPs are extensively studied in biological function owing to the structural property of these membrane-bound proteins invovled in enzymatic reaction. Conventional approaches for CYPs purification need extraction from the membrane into detergent micelles to form detergent-solubilized samples. However, detergent can induce changes on the conformation of CYPs. Stable solubilization and native conformation of CYPs is therefore an important issue in biotechnology as well as in biochemistry and structural biology. Our alternative detergent-free expression system can best preserve the native conformation of CYPs for your specific project.
Figure 1. Schematic of the binding of CO to the heme moiety of CYP. (J. Clin. Biochem. Nutr., 2008)
Creative Biostructure has great expertise in high-yield CYPs production using detergent-free membrane protein expression systmes, we can carry out various strategies for Mempro™ detergent-free CYPs production, including:
Nanodiscs are the self-assembly nanotechnology to stabilize membran proteins removed from the membrane by membrane scaffold proteins (MSPs) in no need of detergent. Creative Biostructure can reconstitute CYPs into nanodiscs in detergent-free methodology: Expressing the CYPs in a cell-free system with the addition of pre-assembled nanodiscs.
The amphipathic amphipols have the ability to “trap” around transmembrane regions of proteins, allowing them to stay folded. Creative Biostructure can apply A8-35 to solubilize CYPs during purification step.
The SMALPs are self-assembled by the simple addition of the SMA co-polymer. At neutral or alkaline pH, a disc-like structure assembles itself, encapsulating cytochrome P450 in a form amenable to be purified.
These novel detergent-free purification approaches for cytochrome P450 production can be obtained easily, and facilitates more comprehensively structural and functional studies.
Creative Biostructure can provide various custom Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
References:A. Nakao, et al. (2008). Therapeutic antioxidant medical gas. J. Clin. Biochem. Nutr., 44(1): 1-13.C. Tribet, et al. (1996). Amphipols: Polymers that keep membrane proteins soluble in aqueous solutions. Proc Natl Acad Sci U. S. A. 93: 15047-15050.M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans. 39: 813-818.K. S. Bhat and G. Padmanaban (1978). Cytochrome P450 synthesis in vivo and in a cell-free system from rat liver. FEBS Lett., 89(2): 337-340.