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Mempro™ Cytochrome P450 Production Using Virus-Like Particles System

Creative Biostructure can offer advanced custom Mempro™ cytochrome P450 (CYP) production services using virus-like particles system. You can count on us all through your projects.

MemproTM Cytochrome P450 Production Using Virus-Like Particles System Figure 1. Structures of CYP158A1 and CYP158A2.

Virus-like particles (VLPs) can mimic the native virus, are however non-infectious owing to they do not have any viral genetic materials. VLPs are self-assembly multiprotein structures, which are widely used in the field of vaccinology. Up to now, VLPs have been generated from components of a wide variety of virus families including bacteriophages (e.g. Qβ, AP205), Flaviviridae (e.g. Hepatitis C virus), Retroviridae (e.g. HIV), and Parvoviridae (e.g. adeno-associated virus). 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, including G protein-coupled receptors (GPCRs), ion channels. Cytochromes P450 are a superfamily of hemoproteins containing a hemo-iron center. Human CYPs are membrane-associated proteins located in the inner membrane of mitochondria or in the endoplasmic reticulum of cells. CYPs are the major enzymes invovled in drug metabolism.

VLPs can be performed for a wide variety of applications, including:

  • Antibody screening;
  • Phage and yeast display;
  • Immunogens/vaccines production;
  • Ligand binding assays;
  • Nucleic acids and small molecules delivery.
  • Other potential applications

Creative Biostructure provides high-yield CYPs in the stable, highly purified and native-conformation state using VLPs system. Lipoparticles can be produced from bacterial cells, yeast cells, insect cells, plant cells and mammalian cells for rhodopsin-like receptors and pumps production. Well-characterized commercial 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, 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 CYPs within lipoparticles are derived directly from the cell surface without mechanical disruption or detergents, the native structure and orientation of CYPs are preserved.

Creative Biostructure provides other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
D. C. Lamb and M. R. Waterman. (2013). Unusual properties of the cytochrome P450 superfamily. Philos. Trans. R. Soc. Lond B Biol. Sci., 368(612): 20120434.
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.
S. Willis, et al. (2008). Virus-like particles as quantitative probes of membrane protein interactions. Biochemistry, 47(27): 6988-6890.
Virus-like particles. (https://en.wikipedia.org/wiki/Virus-like_particle#Assembly_of_VLPs).


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