Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure can offer custom Mempro™ alpha/beta-hydrolase production services using virus-like particles.
Virus-like particles (VLPs) can mimic the native virus, but are non-infectious owing to they do not have any viral genetic materials. VLPs are seclf-assembly multiprotein structures, which are widely used in the field of vaccinology. VLPs are mostly derived from the Hepatitis B virus and composed of the small HBV derived surface antigen (HBsAg). Recently, virus-like particles carrying conformationally-complex membrane proteins (termed lipoparticles) have been applied for integral membrane protein production. Lipoparticles can incorporate a wide range of structurally intact membrane proteins, such as G-protein coupled receptors (GPCRs). Members of alpha/beta-hydrolase superfamily belong to a large class of hydrolytic enzymes, which possess an alpha/beta-hydrolase fold containing 8 beta strands connected by 6 alpha helices. A wide range of enzymes, such as proteases, lipases, esterases, dehalogenases and epoxide hydrolases, have the alpha/beta-hydrolase fold structure.
Figure 1. Crystal structure of a putative alpha/beta hydrolase from Nostoc sp PCC 7120.
Virus-like particles can be performed for a wide range of applications, including:
Creative Biostructure can provide high-yield alpha/beta-hydrolases in the stable, highly purified and native-conformation state. Lipoparticles can be produced from bacterial cells, yeast cells, insect cells, plant cells and mammalian cells for alpha/beta-hydrolase production. Escherichia coli (E. coli) strains, insect cells and mammalian cells are the most widely used systems for VLP production. For instance, we can obtain lipoparticles from mammalian cells by co-expressing the retroviral structural core polyprotein, Gag, along with a desired membrane protein. Gag core proteins self-assemble at the plasma membrane, where they bud off and capture target membrane proteins. Since the alpha/beta-hydrolases within lipoparticles are derived directly from the cell surface without mechanical disruption or detergents, the native structure and orientation of alpha/beta-hydrolases are maintained.
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). A. Roldão, et al. (2010). Virus-like particles in vaccine development. Expert Rev. Vaccines, 9(10): 1149-1176.M. Holmquist (2000). Alpha/beta-hydrolase fold enzymes: structures, functions and mechanisms. Curr. Protein Pept. Sci., 1(2): 209-35. N. Lenfant, et al. (2012). ESTHER, the database of the α/β-hydrolase fold superfamily of proteins: tools to explore diversity of functions. Nucl. Acids. Res., 41: D423-429.S. Willis, et al. (2008). Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry, 47(27): 6988-6890.