Creative Biostructure can offer advanced custom Mempro™ protein production services for peptidase MA using cell-free expression system. Our patent cell-free expression systems can best preserve the native conformation of peptidase MA.
Mempro™ cell-free protein production system is an innovative platform to produce membrane proteins, which can bypass the limitations that encountered in cell-based protein production system, including host cell toxicity, low yield and solubilization and purification using detergents. Peptidase MA is actually a kind of zinc-dependent metallopeptidase, of which a divalent cation (such as zinc) can activate the water molecule. This metal ion is held in place by amino acid ligands, usually three in number. In some cases of co-catalytic metallopeptidases, two metal ions can be observed in crystallography ligated by five amino acids, with one amino acid ligating both metal ions. Additionally, the known metal ligands are usually His, Glu, Asp or Lys. Peptidase MA is one of the two zinc-dependent metallopeptidases, containing the HEXXH motif with the two His residues as zinc ligands.
Figure 1. The structural model of stromelysin-1 (OPM Database)
Creative Biostructure has a wealth of experience in high-yield peptidase MA production using cell-free protein 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 widely employed for peptidase MA production. Wheat germ extract can provide eukaryotic translation system for peptidase MA expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for peptidase MA 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 employed for eukaryotic cell-free transmembrane 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.
With the Mempro™ cell-free protein production platform, Creative Biostructure is capable of expressing, purifying and crystallizing peptidase MA to facilitate the study of their biological functions.
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 peptidase MA 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. Gaczynska, et al. (1994). Peptidase activities of proteasomes are differentially regulated by the major histocompatibility complex-encoded genes for LMP2 and LMP7. Proc. Nat. Acad. Sci. U. S. A., 91(20): 9213-9217. Peptidase M10, metallopeptidase. InterPro. (http://www.ebi.ac.uk/interpro/entry/IPR001818)