Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure provides specific costum Mempro™ protein production services for galactose-binding domain-like proteins based on cell-free expression system.
Mempro™ cell-free protein production system is an innovated system to produce membrane proteins, which bypasses the limitations that encountered in cell-based production system, such as host cell toxicity, low yield and solubilization and purification using detergents. Galactose-binding domain-like proteins are composed of a beta-sandwich, in which the beta-strands can form the beta-sheets. Proteins possessing a galactose-binding domain-like fold can be found in both eukaryotes and prokaryotes. The common function of galactose-binding domains is to bind carbohydrate and have carbon-oxygen lyase activity, such as phospholipids on the outer side of the mammalian cell membrane for coagulation factor Va, and membrane-anchored ephrin for the Eph family of receptor tyrosine kinases. It has proven that the galactose-binding domain-like proteins play an important role in carbohydrate metabolic process.
Figure 1. The structural model of SUN domain-containing protein 2. (OPM database)
Creative Biostructure has extensive expertise in high-yieldgalactose-binding domain-like protein production using cell-free protein expression system, we can perform various strategies for Mempro™ cell-free protein production, including:
Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is widely used for galactose-binding domain-like protein production. Wheat germ extract can provide eukaryotic translation system for galactose-binding domain-like protein expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for galactose-binding domain-like protein expression. Rabbit reticulocyte lysate is a powerful tool for eukaryotic cell-free membrane protein expression. To employ an appropriate source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered. In addition, one challenge associated with MemproTM cell-free membrane protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize MemproTM cell-free protein production for galactose-binding domain-like proteins using plasmids due to they are circular and have no end for the exonucleases to attach.
With the Mempro™ cell-free protein production platform, Creative Biostructure are capable of expressing, purifying and crystallizing galactose-binding domain-like 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.Galactose-binding domain-like (IPR008979). (http://www.ebi.ac.uk/interpro/entry/IPR008979)Galactose-binding domain-like protein. (http://www.uniprot.org/uniprot/K2RZ34)