Sortase A-Mediated N-Terminal Modification of Cowpea Chlorotic Mottle Virus for Highly Efficient Cargo Loading.
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.
Radboud University , Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
Lise
L
Schoonen
Radboud University , Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
Jan
J
Pille
Radboud University , Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
Annika
A
Borrmann
Radboud University , Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
Roeland J M
RJ
Nolte
Radboud University , Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
Jan C M
JC
van Hest
D019881:Aminoacyltransferases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D017795:Bromovirus / Q000737:chemistry*; D002213:Capsid / Q000737:chemistry*; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D008958:Models, Molecular; D011506:Proteins / Q000008:administration & dosage / Q000737:chemistry; D054852:Small Molecule Libraries / Q000008:administration & dosage / Q000737:chemistry
D001426:Bacterial Proteins; D004337:Drug Carriers; D011506:Proteins; D054852:Small Molecule Libraries; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases