Creative Biostructure provides incomparable custom Mempro™ phox homology (PX) domain-containing production services using detergent-free expression system.
PX domain is a conserved phospholipid-binding domain that are composed of approximately 120 amino acids. For example, PX domain protein can bind specifically phosphatidylinositol 3-phosphate, also termed PtdIns(3)P. PX domain functions as crucial roles in membrane trafficking, cell signaling, lipid modification and protein sorting. With conventional approaches, detergent is required for PX domain protein production by extraction from the membrane. However, detergent can induce changes on membrane protein conformation. Stable solubilization and native conformation of PX domain-containing protein are therefore important issues in biotechnological field. Our alternative Mempro™ detergent-free expression system can best preseve the native conformation of PX domain proteins for further structural studies.
Figure 1. Schematics of protein sorting assisted by PX domain proteins. (Nature, 2005)
Creative Biostructure have great expertise in high-yield PX domain-containing protein production using detergent-free membrane protein expression system, we can perform various strategies for Mempro™ detergent-free PX-domain protein production, including:
Nanodiscs are the self-assembly nanotechnology to stabilize membrane proteins removed from the membrane by membrane scaffold proteins (MSPs) in no need of detergent. Creative Biostructure can reconstitute PX-domain proteins into nanodiscs in detergent-free methodology: Expressing the desired membrane protein in the cell-free system with the addition of pre-assembled nanodiscs.
The amphipathic amphipols have the ability to “trap” around membrane proteins of interest, allowing them to stay folded. Creative Biostructure can apply A8-35 to solubilize PX-domain protein during purification process.
The SMALPs are self-assembled by the simple addition of the SMA co-polymers. At neutral or alkaline pH, a disc-like structure assembles itself, encapsulating the PX-domain protein in a form amenable to be purified.
These novel detergent-free technologies for PX domain containing protein production can be obtained easily, and 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:C. S. Vollert and P. Uetz (2004). The phox homology (PX) domain protein interaction network in yeast. Mol. Cell Proteomics., 3(11):1053-1064.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.H. T. McMahon and J. L. Gallop (2005). Membrane curvature and mechanisms of dynamic cell membrane remodelling. Nature, 438: 590-596.L. F. Seet and W. Hong (2006). The Phox (PX) domain proteins and membrane traffic. Biochim. Biophy. Acta, 1761(8): 878-896. M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans., 39: 813-818. PX domain. (https://en.wikipedia.org/wiki/PX_domain)