With the most advanced technology and years of experiences in antibody engineering field, Creative Biostructure now provide specific custom Mempro™ alpha/beta-hydrolase production services using detergent-free expression system.
Alpha/beta-hydrolase containing an alpha/beta-hydrolase fold structure is a group of hydrolytic enzymes with diverse catalytic functions. Alpha/beta-hydrolase fold has an alpha/beta sheet motif composed of 8 beta-strands connected by 6 alpha-helices. Proteins containing this alpha/beta-hydrolase fold do not share sequence similarity, such as acetylcholinesterase, lipase, dienelactone hydrolase, proline oligopeptidase, thioesterase, proline iminopeptidase, haloalkane dehalogenase, serine carboxypeptidase, haloperoxidase, epoxide hydrolase, and hydroxynitrile lyase. For most of the family members, the beta-strands are paralleled, but some have an inversion of the first strand, which gives it an antiparallel orientation. Conventional approaches for alpha/beta-hydrolase production require extraction from the membrane into detergent micelles to form detergent-solubilized samples. However, detergent can induce changes on the conformation of proteins. Our substitutive detergent-free expression system can best preserve the native conformation of alpha/beta-hydrolases for further structural analysis.
Figure 1. A model of alpha/beta-hydrolase fold. (Wikipedia)
Creative Biostructure has advanced expertise in high-yield alpha/beta-hydrolase production using detergent-free membrane protein expression system, we can perform various strategies for Mempro™ detergent-free alpha/beta-hydrolase production, including:
Nanodiscs are the self-assembly nanotechnology to stabilize membrane proteins removed from the membrane by membrane scaffold proteins (MSPs) in absence of detergent. Creative Biostructure can reconstitute alpha/beta-hydrolases into nanodiscs in detergent-free methodology: Expressing the alpha/beta-hydrolases in a cell-free system with the addition of pre-assembled nanodiscs.
The amphipathic amphipols have the ability to “trap” over transmembrane regions of proteins, allowing them to stay folded. Creative Biostructure can apply A8-35 to solubilize alpha/beta-hydrolases during purification process.
The SMALPs can be self-assembled by simply adding SMA co-polymers. At neutral or alkaline pH condition, a disc-like structure assembles itself, encapsulating the alpha/beta-hydrolases in a formation amenable to be purified.
These novel detergent-free technologies for membrane proteins 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:Alpha/beta hydrolase fold. (https://en.wikipedia.org/wiki/Alpha/beta_hydrolase_fold). 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.D. L. Ollis, et al. (1992). The α/β hydrolase fold. Protein Eng., 5(3): 197-211.M. Jamshad, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans., 39: 813-818.