Based on the comprehensive protein engineering platform established through years of experience, scientists form Creative Biostructure offers tailored Mempro™ peptidase MA production services using detergent-free expression system.
Peptidase MA is a group of zinc-dependent metallopeptidase, of which a divalent cation (such as zinc) can activate the water molecule. Over fifty metallopeptidase families have been identified. The 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. Conventional approaches for membrane protein purification need extraction from the membrane into detergent micelles. However, detergent can induce conformational change on membrane proteins. Our alternative detergent-free expression system can best preserve the native conformation of peptidase MA for further structural studies.
Figure 1. The structural model of Endoplasmic reticulum aminopeptidase 1, one member of peptidase family M1. (OPM Database)
Creative Biostructure has rich professional experience in high quality peptidase MA production using detergent-free membrane protein expression system, we can perform various strategies for Mempro™ detergent-free protein production, including:
Nanodiscs are the self-assembly system to stabilize membrane proteins removed from the membrane by membrane scaffold proteins (MSPs) in no need of detergent. Creative Biostructure can reconstitute peptidase MA into nanodiscs in detergent-free process: Expressing the peptidase MA in a cell-free system with the addition of pre-assembled nanodiscs.
The amphipathic amphipols have the ability to “trap” around transmembrane regions of proteins, allowing them to stay folded. Creative Biostructure can utilize A8-35 to solubilize peptidase MA during purification step.
The SMALPs are self-assembled by the simple addition of the SMA co-polymers. At neutral or alkaline pH, a disc-like structure assembles itself, encapsulating peptidase MA in a form amenable to be purified.
These novel detergent-free technologies for peptidase MA production can be obtained easily, and enabling more comprehensively 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:C. Tribet, et al. (1996). Amphipols: Polymers that keep membrane proteins soluble in aqueous solutions. Proc Natl Acad Sci U. S. A. 93: 15047-15050. 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.M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans., 39: 813-818.Peptidase M10, metallopeptidase. InterPro. (http://www.ebi.ac.uk/interpro/entry/IPR001818)