Creative Biostructure offers advanced custom Mempro™ protein production services for Eicosanoid and Glutathione metabolism proteins (MAPEG) using cell-free expression system. Our patent cell-free expression systems can best preserve the native conformation of MAPEG (Membrane-Associated Proteins in Eicosanoid and Glutathione metabolism).
MAPEG is a superfamily of membrane protein defined by their structure, enzyme activity and the sequence motif. MAPEG is associated with the biosynthesis of arachidonic-derived mediators of pain, fever, and inflammation and also the bio-transformation and detoxification of electrophilic substances. This superfamily consists of six human proteins and seven members from plant and bacteria. Based on the sequence similarity, two subgroups could be defined. Microsomal glutathione transferase 1 (MGST1) share 38% sequence identity while the second group has less than 20% identity, suggesting two evolutional pathways. Members of MAPEG superfamily have quite divergent functions. Leukotriene C4 (LTC4) synthase is an enzyme that catalyzes the conversion from leukotriene A4 (LTA4) and glutathione to leukotriene C4. There are five α-helices in LTC4 synthase, in which four are transmembrane helices and one is exposed to the solvent. LTC4 synthase monomers form a cavity to bind glutathione. The hydrophobic environment created by the association between the LTC4 synthase trimer and the glutathione facilitates the docking of LTA4 produced by 5-lipoxygenase pathway.
Figure 1. Crystal structure of human leukotriene C4 synthase.
Creative Biostructure has a wealth of experience in high-yield MAPEG production using cell-free protein expression systems, we can provide various strategies for Mempro™ cell-free protein production, including:
Among Mempro™ cell-free membrane protein production systems, crude rabbit reticulocyte lysate is the most widely employed for eukaryotic cell-free transmembrane protein expression. To employ an appropriate source of the extract, many factors should be considered, such as the availability of materials, protein origin and complexity, downstream processing, etc. Crude rabbit reticulocyte lysate is advanced in performing the post-translational modifications on the protein as well as its easy preparation.
With the Mempro™ cell-free protein production platform, Creative Biostructure is capable of expressing, purifying and crystallizing MAPEG to facilitate the study of their biological functions.
We provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
References:F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.Ago H, Kanaoka Y, Irikura D, et al. Crystal structure of a human membrane protein involved in cysteinyl leukotriene biosynthesis[J]. Nature, 2007, 448(7153): 609-612.Hebert H, Jegerschöld C. The structure of membrane associated proteins in eicosanoid and glutathione metabolism as determined by electron crystallography[J]. Current opinion in structural biology, 2007, 17(4): 396-404.