Creative Biostructure provides customized Mempro™ cell-free protein production services for receptor-type kinases, including but not limited to receptor tyrosine kinases, receptor serine kinases and receptor threonine kinases.
MemproTM cell-free protein production system is the innovative systems to produce integral membrane proteins, which can bypass the limitations that encountered in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification using detergents. Receptor-type kinases are a family of membrane-associated receptor proteins for numerous cytokines, hormones, and polypeptide growth factors, which play an important role in signaling transduction. Receptor-type receptors can immediately activate several signaling pathways, such as inositol-phospholipid-calcium pathway and Ras pathway. The kinase activity of is triggered after the interaction between a ligand and receptor-type kinase, then a phosphorylation cascade can transmit the signal. Different kinases enable adding phosphate groups to amino acids that possess a hydroxyl (OH) group containing side chain.
Figure 1. A Model of biological function of receptor tyrosine kinase.
Creative Biostructure has extensive expertise in overexpression, purification and characterization of receptor-type kinases, we can provide various strategies for desired membrane proteins, including:
Among Mempro™ cell-free protein production platform, E. coli S30 extract is the most widely used for receptor-type kinase production. Wheat germ extract can provide eukaryotic translation environment for receptor-type kinase expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major tool for receptor-type kinase production, 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 system. To select an appropriate source for protein expression, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.
Notably, One challenge associated with Mempro™ cell-free protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize this expression system using plasmids owing to they are circular and have no end for the exonucleases to attach.
We can 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.M. A. Lemmon and J. Schlessinger (2010). Cell signaling by receptor-tyrosine kinases. Cell, 141(7): 1117-1134.Receptor tyrosine kinase. (https://en.wikipedia.org/wiki/Receptor_tyrosine_kinase).