Engineering Recombinant Virus-like Nanoparticles from Plants for Cellular Delivery
| Autor: | Garry Morgan, Frank Sainsbury, Philippe V. Jutras, Eva C. Thuenemann, Michael J. Landsberg, Noor H. Dashti, George P. Lomonossoff, Lou Brillault |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Integrins Materials science Cryo-electron microscopy Recombinant Fusion Proteins Integrin General Physics and Astronomy 02 engineering and technology Protein Engineering Recombinant virus law.invention Green fluorescent protein 03 medical and health sciences Drug Delivery Systems law Tobacco Humans Nanotechnology General Materials Science Cloning Molecular Plant Proteins Drug Carriers Bioconjugation biology General Engineering RNA 021001 nanoscience & nanotechnology Molecular biology Plant Leaves 030104 developmental biology Capsid MCF-7 Cells Recombinant DNA Biophysics biology.protein Nanoparticles 0210 nano-technology Bluetongue virus |
| Zdroj: | ACS Nano. 11:3476-3484 |
| ISSN: | 1936-086X 1936-0851 |
| Popis: | Understanding capsid assembly following recombinant expression of viral structural proteins is critical to the design and modification of virus-like nanoparticles for biomedical and nanotechnology applications. Here, we use plant-based transient expression of the Bluetongue virus (BTV) structural proteins, VP3 and VP7, to obtain high yields of empty and green fluorescent protein (GFP)-encapsidating core-like particles (CLPs) from leaves. Single-particle cryo-electron microscopy of both types of particles revealed considerable differences in CLP structure compared to the crystal structure of infection-derived CLPs; in contrast, the two recombinant CLPs have an identical external structure. Using this insight, we exploited the unencumbered pore at the 5-fold axis of symmetry and the absence of encapsidated RNA to label the interior of empty CLPs with a fluorescent bioconjugate. CLPs containing 120 GFP molecules and those containing approximately 150 dye molecules were both shown to bind human integrin via a naturally occurring Arg-Gly-Asp motif found on an exposed loop of the VP7 trimeric spike. Furthermore, fluorescently labeled CLPs were shown to interact with a cell line overexpressing the surface receptor. Thus, BTV CLPs present themselves as a useful tool in targeted cargo delivery. These results highlight the importance of detailed structural analysis of VNPs in validating their molecular organization and the value of such analyses in aiding their design and further modification. |
| Databáze: | OpenAIRE |
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