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Sortase A‑Mediated N‑Terminal Modification of Cowpea Chlorotic Mottle Virus for Highly Efficient Cargo Loading

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journal contribution
posted on 16.12.2015, 00:00 by Lise Schoonen, Jan Pille, Annika Borrmann, Roeland J. M. Nolte, Jan C. M. van Hest
A new strategy is described for the modification of CCMV for loading of cargoes inside the viral capsid. Sortase A, an enzyme which is present in Gram-positive bacteria, was used to attach cargo to the glycine-tagged N-termini of several CCMV variants. We show that small molecules and proteins bearing a C-terminal LPETG-motif can be attached in this way. This method allows for the site-specific, covalent, and orthogonal modification of CCMV capsids in a mild fashion, leading to high encapsulation efficiencies. This strategy can easily be expanded to other types of cargoes, labeled with an LPETG-tag without altering protein function.