Mempro™ Cell-Free Bcl-2 Inhibitors of Programmed Cell Death Production

Creative Biostructure can provide costum Mempro™ protein production services for Bcl-2 inhibitors of programmed cell death based on cell-free expression system.

Mempro™ cell-free protein production system is the innovated system to produce membrane proteins, which overcome the difficulties that encountered in cell-based production system, such as host cell toxicity, low yield and solubilization and purification using detergents. B-cell lymphoma 2 (Bcl-2) is a family of evolutionarily related proteins that function as apoptosis regulators (programmed cell death). The Bcl-2 family share a structural domoin that consists of a hydrophobic helix surrounded by amphipathic helices. Most of members in Bcl-2 family have transmembrane domains. It is reproted that the pro-survival Bcl-2-like proteins (Bcl-2, Bcl-xL, Bcl-w, Mcl-1, and A1) bind to and inhibit Bax and Bak, which are the terminal effectors directly mediate mitochondrial outer membrane permeabilization. Additionally, Bcl-2 inhibitors of programmed cell death superfamily proteins can interact in an inhibitory manner with BCL-2, BCL-XL, and BCL-w.

Figure 1. The structural model of human Bcl-xL, an inhibitor of programmed cell death. (Nature, 1996)

Creative Biostructure has extensive expertise in high-yield Bcl-2 inhibots superfamily protein production using cell-free protein expression system, 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 widely used for Bcl-2 inhibitors superfamily protein production. Wheat germ extract can provide eukaryotic translation system for Bcl-2 inhibitors superfamily protein expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for Bcl-2 inhibitors superfamily protein expression. Rabbit reticulocyte lysate is an important tool for eukaryotic cell-free transmembrane protein expression. 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 are capable of expressing, purifying and crystallizing Bcl-2 inhibitors superfamily proteins to perform further 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:
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.
S. Cang, et al. (2015). ABT-199 (venetoclax) and BCL-2 inhibitors in clinical development. J. Hematol. Oncol., 8: 129.
S. W. Muchmore, et al. (1996). X-ray and NMR structure of human Bcl-xL, an inhibitor of programmed cell death. Nature, 381(6580): 335-341.