Artificial Protein Cage Delivers Active Protein Cargos to the Cell Interior.

Antonina Naskalska,Izabela Stupka,Jonathan G Heddle,Michał Bochenek,Jan Różycki,Kinga Borzęcka-Solarz,Elżbieta Pyza

doi:10.1021/acs.biomac.1c00630

Antonina Naskalska, Izabela Stupka + Show 5 more

Open Access

https://doi.org/10.1021/acs.biomac.1c00630

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Abstract

Artificial protein cages have potential as programmable, protective carriers of fragile macromolecules to cells. While natural cages and VLPs have been extensively exploited, the use of artificial cages to deliver active proteins to cells has not yet been shown. TRAP-cage is an artificial protein cage with an unusual geometry and extremely high stability, which can be triggered to break apart in the presence of cellular reducing agents. Here, we demonstrate that TRAP-cage can be filled with a protein cargo and decorated with a cell-penetrating peptide, allowing it to enter cells. Tracking of both the TRAP-cage and the cargo shows that the protein of interest can be successfully delivered intracellularly in the active form. These results provide a valuable proof of concept for the further development of TRAP-cage as a delivery platform.

Highlights

Transport of molecular cargos to cells is desirable for a range of applications including delivery of drugs, genetic material, or enzymes
Artificial cages allow other features such as geometries not seen in natural cages,[8] which widens the possible library of building blocks, and replacement of enhancement of protein− protein interactions with other interactions, leading to control of disassembly.[8,10]
We show for the first time that an artificial protein cage is capable of delivering a functional protein cargo to the cell interior

Summary

Introduction

Transport of molecular cargos to cells is desirable for a range of applications including delivery of drugs, genetic material, or enzymes. A number of nanoparticles have been employed to achieve this including liposomes,[1] virus-like particles,[2] nonviral protein cages,[3] DNA origami cages,[4,5] and inorganic nanoparticles.[6] Protein cages are a promising approach as demonstrated by viruses in nature which are able to deliver genetic material to cells, often with high effectiveness and specificity. The advantage of using such an approach is that the resulting cages can be given properties and capabilities that may not be available or feasible in naturally occurring forms. This includes triggerable assembly,[8,9] which allows substituent proteins to be expressed as relatively small individual subunits with/without cargos, circumventing possible production problems associated with large complex formation in the cell. We show for the first time that an artificial protein cage is capable of delivering a functional protein cargo to the cell interior

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