|Maize Rayado Fino Virus-VLPs and peptide crosslinking|
Maize Rayado Virus-VLPs and peptide crosslinking on the viral surface
Figure legend: Two step protein cross-linking procedures between Cys 2-VLPs and F and HN peptides. (A) Cross-linking of NHS-PEG4-Maleimide with amine groups (N-terminal) of F and HN peptides. (B) Cross-linking of peptide-NHS-PEG4-Maleimide with thiol group Cys 2-VLPs.
Natilla, A,Hammond RW, Maize rayado fino virus-like particles expressed in tobacco plants: a new platform for cysteine selective bioconjugation peptide display -J Virol Methods. 2011 Dec;178(1-2):209-15. doi: 10.1016/
Maize rayado fino virus (MRFV) (genus Marafivirus, family Tymoviridae) iscomposed of isometric particles of 30 nm in diameter. Each particle has 180 copies of the capsid protein (CP) forming a T=3 structure, stabilized by protein-protein interactions. MRFV CPs self-assemble in Escherichia coli and in Nicotiana benthamianainto virus-like particles (VLPs) identical in size and shape to MRFV native capsids, a very desirable attribute in nanotechnology (Hammond and Hammond, 2010). Our laboratory investigated MRFV- VLPs produced in tobacco plants for their ability to serve as platform to which different peptides could be covalently displayed.
To provide an anchor for chemical modifications, we created Cys-MRFV-VLPs mutants by substituting several of the amino acids present on the shell of the wild-type MRFV-VLPs with cysteine residues. The mutant designated Cys 2-VLPs exhibited, under native conditions, cysteine thiol reactivity in bioconjugation reactions with a fluorescent dye. Successively, Cys 2-VLPs was cross-linked by NHS-PEG4-Maleimide to 17 (F) and 8 (HN) amino acid long peptides, corresponding to neutralizing epitopes of Newcastle disease virus(NDV). The resulting Cys 2-VLPs-F and Cys 2-VLPs-HN were recognized in Western blots by antibodies to MRFV as well as to F and HN.
The results demonstrated that plant-produced MRFV-VLPs have the ability to function as a novel platform for the multivalent display of surface ligands.