Creative Biostructure has developed the first-class membrane protein production platform to provide custom lipoxygenase production service using virus-like particles expression system.
Lipoxygenase is a member of arachidonate cascade enzymes that are ubiquitous in many animals and plants. It catalyzes the dioxygenation of polyunsaturated fatty acids for the production of fatty acid hydroperoxides. A lipoxygenase molecule contains one iron atom in its active site, which is critical for lipoxygenase's catalysis activity. Lipoxygenases can be categorized into 4 types based on their regiospecificity: arachidonate 5-, 8-, 12- and 15- lipoxygenase (5-LOX, 8-LOX, 12-LOX and 15-LOX). These lipoxygenases catalyze the dioxygenation on carbon 5, 8, 12 and 15 of arachidonate acid. In plants, lipoxygenases are involved in many aspects such as growth and development, response to wounding, pest resistance, etc. In mammalians, lipoxygenases are involved in xenobiotic metabolism.
Figure 1. The schematic structure of Arachidonate 15-lipoxygenase. (OPM Database)
Virus-like particles (VLPs) are self-assembled, highly organized structural subunits defined as viral "empty shells" which are assembled from viral structural proteins. VLPs mimic the conformation and organization of native virus, but all of the VLPs are devoid of any viral genetic materials. Various host systems have been successfully developed to obtain VLPs. VLPs have been widely used in vaccine development and production programs, antigen deliveries, heterologous epitope presentation, and recombinant membrane protein productions. VLPs and lipoparticles have been reported to be applied for various structurally intact membrane protein productions successfully, including ion channels, G protein-coupled receptors (GPCRs), etc.
Creative Biostructure provides stable, high-yield and high-purity lipoxygenase production service using the most advanced VLP technique. With years of experience, our science team has successfully developed multiple VLP host systems to meet different experimental requirements. These host systems including but not limited to:
Bacteria (E. coli) YeastGreen plants (tobacco, alfalfa, potato, etc.)Insect cellsMammalian cells
E. coli and insect cells are the most widely applied systems for VLP production. Mammalian cells are also used for VLP productions in vaccine developments and gene therapy programs. Lipoparticles will be obtained by co-expressing the retroviral structural core polyprotein, Gag, along with the target lipoxygenase in mammalian cells. Gag core proteins perform self-assembling at the plasma, and bud-off from the plasma membrane as lipoparticles (VLPs). The expressed lipoxygenase will be captured by the lipoparticles, and harvested within the lipoparticles from the surfaces of cells. In addition, native lipoxygenase can be isolated without detergents or mechanical disruption, so its orientation of membrane proteins and the native structure will be retained.
Creative Biostructure also provides other Mempro™ membrane protein production services with multiple technologies. Welcome to contact us for more details.
References:Feussner I, Wasternack C. The lipoxygenase pathway [J]. Annual review of plant biology, 2002, 53(1): 275-297.A.Roldão, et al. (2010). Virus-like particles in vaccine development.Expert Rev. Vaccines, 9(10): 1149-1176.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.