Predicting the loading of virus-like particles with fluorescent proteins

Biomacromolecules
W Frederik RurupJeroen J L M Cornelissen

Abstract

The virus-like particle (VLP) of the Cowpea Chlorotic Mottle Virus (CCMV) has often been used to encapsulate foreign cargo. Here we show two different rational design approaches, covalent and noncovalent, for loading teal fluorescent proteins (TFP) into the VLP. The covalent loading approach allows us to gain control over capsid loading on a molecular level. The achieved loading control is used to accurately predict the loading of cargo into CCMV VLP. The effects of molecular confinement were compared for the differently loaded VLPs created with the covalent method. We see that the loading of more than 10 fluorescent proteins in the 18 nm internal cavity of the CCMV capsid gives rise to a maximum efficiency of homo-FRET between the loaded proteins, as measured by fluorescence anisotropy. This shows that already at low levels of VLP loading molecular crowding starts to play a role.

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Citations

Feb 18, 2014·Journal of the American Chemical Society·W Frederik RurupJeroen J L M Cornelissen
Dec 5, 2015·Journal of the American Chemical Society·Reinhard Zschoche, Donald Hilvert
Dec 24, 2015·Angewandte Chemie·Yusuke AzumaDonald Hilvert
Dec 30, 2015·Nanomedicine : Nanotechnology, Biology, and Medicine·Ding GaoZong-Qiang Cui
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Jan 6, 2017·Journal of the American Chemical Society·Melanie BraschJeroen J L M Cornelissen
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