Mempro™ Cell-Free Rhodopsin Production

Creative Biostructure provides customized Mempro™ protein production services for rhodopsin-like receptors and pumps using cell-free expression system. Our cell-free expression systems can best preserve the native activity of rhodopsin-like receptors and pumps. You can count on us all through your projects.

Mempro™ cell-free protein production systems are the innovative systems to produce G-protein coupled receptors (GPCRs), which can bypass the limitations that encountered in cell-based protein production systems, such as host cell toxicity, low yield and solubilization and purification using detergents (Figure 1). Rhodopsin-like receptors and pumps are a family of proteins that comprise the largest group of GPCRs. The rhodopsin-like receptors and pumps transduce extracellular signals through interaction with guanine nucleotide-binding proteins.

Figure 1. Cell-free (CF) expression models for membrane protein production. P-CF indicates synthesis of membrane proteins in the absence of supplied hydrophobic compartments and precipitate after translation. D-CF indicates membrane proteins that kept in the soluble form by insertion into provided detergent micelles. L-CF indicates membrane proteins can be inserted into preformed liposomes, bicelles or nanodiscs. (http://www.sfb807.de/volker-doetsch-frank-bernhard.html)

Creative Biostructure has a wealth of experience in high-yield rhodopsin-like receptors and pumps production using cell-free protein expression systems, we can provide various strategies for Mempro™ cell-free protein production, including:

• Mempro™ Cell-Free Protein Production in E. coli;
• Mempro™ Cell-Free Protein Production in Wheat Germ;
• Mempro™ Cell-Free Protein Production in Rabbit Reticulocyte;
• Mempro™ Cell-Free Protein Production in Baculovirus.

Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is commonly used for rhodopsin-like receptors and pumps production. Wheat germ extract can provide eukaryotic translation system for rhodopsin-like receptors and pumps expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for rhodopsin-like receptors and pumps expression, which is developed in the early stage that transfected insect cells in combination with vectors derived from the baculovirus. Crude rabbit reticulocyte lysate is also the most widely used for eukaryotic cell-free membrane protein expression systems. To select an appropriate source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.

With the Mempro™ cell-free protein production platform, Creative Biostructure is capable of expressing, purifying and crystallizing rhodopsin-like receptors and pumps to facilitate the study of their biological functions. Mempro™ cell-free membrane protein production has a large of advantages, including：

• Very high yield;
• Low cost in time and money;
• Lack of toxicity;
• Incorporation of non-natural amino acids into protein structure;
• Relatively tolerant of additives.

Most notably, One challenge associated with Mempro™ cell-free membrane protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize Mempro™ cell-free membrane protein production for rhodopsin-like receptors and pumps using plasmids due to they are circular and have no end for the exonucleases to attach.

We provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
Cell-free protein synthesis. (https://en.wikipedia.org/wiki/Cell-free_protein_synthesis)
F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.
P2 Cell-free expression of GPCRs: In vitro characterization of the endothelin receptor signal perception and transduction. (http://www.sfb807.de/volker-doetsch-frank-bernhard.html).