Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure provides custom Mempro™ C1 domain-containing protein production services using virus-like particles system.
Virus-like particles (VLPs) can simulate the native virus, but are non-infectious due to they have no viral genetic materials. VLPs are seclf-assembly multiprotein structures, which are popularly used in the field of vaccinology. 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) and ion channels. C1 domains are known as phorbol esters/diacylglycerol binding domains, which are cysteine-rich domains. C1 domains can bind an important sencondary messenger diacylglycerol (DAG) and the analogues of DAG named phorbol esters, which can directly stimulate protein kinase C (PKC). Most notably, C1 domains are composed of about 50 amino acids and are involved in the recruitment of proteins to the membrane.
Figure 1. C1 domain of protein kinase C gamma. (Wikpedia)
Virus-like particles can be applied for a wide range of applications, including:
Creative Biostructure can provide high-yield C1 domain-containing proteins in stable, highly-purified and native-conformation state. Lipoparticles can be generated from bacterial cells, yeast cells, insect cells, plant cells and mammalian cells for C1 domain-containing protein production. Escherichia coli (E. coli) strains, insect cells and mammalian cells are the most widely used tools 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 C1 domain-containing proteins within lipoparticles are derived directly from the cell membrane without mechanical disruption or detergents, the native structure and orientation of C1 domain-containing proteins are retained.
Creative Biostructure provides other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
References:A. Roldão, et al. (2010). Virus-like particles in vaccine development. Expert Rev. Vaccines, 9(10): 1149-1176.F. Colón-González and M. G. Kazanietz (2006). C1 domains exposed: from diacylglycerol binding to protein-protein interactions. Biochim. Biophys. Acta., 1761(8): 827-837. J. Das and G. M. Rahman (2014). C1 domains: structure and ligand-binding properties. Chem. Rev., 114 (24): 12108–12131. S. Willis, et al. (2008). Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry, 47(27): 6988-6890.