Creative Biostructure can provide custom Mempro™ galactose-binding domain-like protein production services using virus-like particles (VLPs).
Galactose-binding domain is composed of a beta-sandwich, in which the strands making up the beta-sheets. There is a high degree of similarity in the beta-sandwich, although an often low level of sequence similarity. It is reported that galactose-binding domain-like proteins can be found in both eukaryotes and prokaryotes. The common function of galactose-binding domains is to bind carbohydrate and have carbon-oxygen lyase activity, such as cell-surface-attached carbohydrate substrates for galactose oxidase and sialidase, and membrane-anchored ephrin for the Eph family of receptor tyrosine kinases. The galactose-binding domain-like proteins play a crucial role in carbohydrate metabolic process.
Figure 1. C2 domain of coagulation factor V. (OPM database)
VLPs are self-assembled multisubunit protein structures, which can mimic the native virus, but are non-infectious due to lacking any viral genetic information. VLPs derived from the Hepatitis B virus and composed of the small HBV derived surface antigen (HBsAg). In addition, VLPs carrying native-conformation membrane proteins (termed lipoparticles) have been employed for the production of integral membrane protein. Lipoparticles can incorporate a wide range of structurally intact membrane proteins, including G protein-coupled receptors (GPCRs), and ion channels.
Creative Biostructure can produce high-yield galactose-binding domain-like proteins 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 galactose-binding domain-like protein production. Escherichia coli (E. coli) strains and insect cells are the most widely used systems for VLPs production. Mammalian cells are also widely used for VLPs production with the target to construct vaccine candidates and gene therapy agents. For instance, lipoparticles can be obtained from mammalian cells by co-expressing the retroviral structural core polyprotein, Gag, along with the membrane protein of interest. Gag core proteins self-assemble at the plasma membrane, where they bud off and capture target membrane proteins. Since the galactose-binding domain-like proteins within lipoparticles are derived directly from the cell surface without mechanical disruption or detergents, the native structure and orientation of galactose-binding domain-like proteins are retained.
Creative Biostructure can also provide 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. D. P. Patterson, et al. (2012). Virus-like particle nanoreactors: programmed encapsulation of the thermostable CelB glycosidase inside the P22 capsid. Soft Matter, 8: 10158-10166. Galactose-binding domain-like (IPR008979). (http://www.ebi.ac.uk/interpro/entry/IPR008979)Galactose-binding domain-like protein. (http://www.uniprot.org/uniprot/K2RZ34) S. Willis, et al. (2008). Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry, 47(27): 6988-6890.