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Mempro™ Cell-Free PH Domain-Like Protein Production

Creative Biostructure provides customized Mempro™ protein production services for pleckstrin homology (PH) domain-like proteins using cell-free expression system. Our cell-free expression systems can best preserve the native activity of PH domain-like superfamily proteins.

Mempro™ cell-free protein production systems are the innovative systems to produce a wide range of integral membrane proteins (including PH domain-like superfamily proteins), 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). PH domains are the small molecular domains that can bind phosphoinositides with high affinity and specificity. The significant property of PH domain is that this specific domain can introduce phosphorylation at different sites within the inositol ringbind lipid. In other words, PH domains have lipid binding specificity.

MemproTM Cell-Free PH Domain-Like Protein Production Figure 1. The schematic of PH domain-like proteins invovled biological function.

Creative Biostructure has extensive expertise in high-yield PH domain-like protein production using cell-free expression systems, we can provide various strategies for Mempro™ cell-free protein production, including:

Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is commonly used for integral membrane proteins, especially PH domain-like proteins. Wheat germ extract can provide eukaryotic translation system for PH domain-like protein expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for PH domain-like protein expression, which is developed in the early stage that cultivated insect cells in combination with the baculovirus vector (bacmid). Crude rabbit reticulocyte lysate is also the most widely used for eukaryotic cell-free membrane protein expression systems. To select the source of the extract, many factors including 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 PH domain-like proteins 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 PH domain-like proteins 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.
M. A. Lemmon (2007). Pleckstrin homology (PH) domains and phosphoinositides. Biochem. Soc. Symp., 74: 81-93.


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