Selective and Efficient Functionalization of P22 Virus-Like Particles Using an Asparaginyl Ligase.
- 1Institute for Molecular Bioscience, Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Queensland, Brisbane, Queensland4072, Australia.
- 2Australian Institute for Bioengineering and Nanotechnology, ARC Research Hub for Advanced Manufacture of Targeted Radiopharmaceuticals, The University of Queensland, Brisbane, Queensland4072, Australia.
- 3Centre for Cell Factories and Biopolymers, Institute for Biomedicine and Glycomics, Australian Research Council Centre of Excellence in Synthetic Biology, Griffith University, Nathan, Queensland4111, Australia.
Abstract
Protein cages provide useful scaffolds for nanoscale engineering because of their highly ordered structures and in vivo self-assembly. These scaffolds are amenable to late-stage conjugation, enabling an expansion in functionality. However, many conjugation techniques either lack site selectivity, require unnatural amino acid incorporation, or have bulky recognition motifs to facilitate ligation reactions. Here, an asparaginyl endopeptidase (AEP) enzyme with ligase activity is employed for the highly efficient functionalization of virus-like particles (VLPs) from Enterobacteria phage P22. The capacity of this enzyme to conjugate peptides and proteins onto assembled P22 VLPs under mild reaction conditions via a minimal extension to the P22 coat protein C-terminus is demonstrated. We extend the reaction efficiency to facilitate a one-pot dual-functionalization reaction, in which two therapeutically relevant receptor-targeting domains are conjugated to P22 VLPs in a single step. Finally, we demonstrate the potential for AEP-mediated bioconjugation to bestow P22 VLPs with receptor-binding functionality in vitro. This work demonstrates the efficacy of AEP ligases as bioconjugation tools for the site-selective functionalization of large molecular assemblies such as VLPs.
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BibTeX
@article{harding2026selective,
title = {{Selective and Efficient Functionalization of P22 Virus-Like Particles Using an Asparaginyl Ligase.}},
author = {Maxim D Harding and Mark A Jackson and Kuok Yap and Pie Huda and David J Craik and Frank Sainsbury and Nicole Lawrence},
journal = {Bioconjugate chemistry},
year = {2026},
month = sep,
doi = {10.1021/acs.bioconjchem.6c00356},
url = {https://europepmc.org/article/MED/42915466},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Selective and Efficient Functionalization of P22 Virus-Like Particles Using an Asparaginyl Ligase.
AU - Maxim D Harding
AU - Mark A Jackson
AU - Kuok Yap
AU - Pie Huda
AU - David J Craik
AU - Frank Sainsbury
AU - Nicole Lawrence
PY - 2026
DA - 2026/09/01
JO - Bioconjugate chemistry
DO - 10.1021/acs.bioconjchem.6c00356
UR - https://europepmc.org/article/MED/42915466
AB - Protein cages provide useful scaffolds for nanoscale engineering because of their highly ordered structures and in vivo self-assembly. These scaffolds are amenable to late-stage conjugation, enabling an expansion in functionality. However, many conjugation techniques either lack site selectivity, require unnatural amino acid incorporation, or have bulky recognition motifs to facilitate ligation reactions. Here, an asparaginyl endopeptidase (AEP) enzyme with ligase activity is employed for the highly efficient functionalization of virus-like particles (VLPs) from Enterobacteria phage P22. The capacity of this enzyme to conjugate peptides and proteins onto assembled P22 VLPs under mild reaction conditions via a minimal extension to the P22 coat protein C-terminus is demonstrated. We extend the reaction efficiency to facilitate a one-pot dual-functionalization reaction, in which two therapeutically relevant receptor-targeting domains are conjugated to P22 VLPs in a single step. Finally, we demonstrate the potential for AEP-mediated bioconjugation to bestow P22 VLPs with receptor-binding functionality in vitro. This work demonstrates the efficacy of AEP ligases as bioconjugation tools for the site-selective functionalization of large molecular assemblies such as VLPs.
SN - 1043-1802
SN - 1520-4812
ER -