Virus-Like Particles (VLPs) as a Platform for Hierarchical Compartmentalization

Area of Science:

• Biomimetic materials science
• Synthetic biology
• Protein nanotechnology

Background:

• Hierarchically self-assembled structures are prevalent in biological systems.
• Designing synthetic analogs that mimic cellular complexity remains a challenge.
• The cell's multicompartmentalized organization inspires the development of advanced functional materials.

Purpose of the Study:

• To design and construct a nested protein cage system for creating synthetic cell-like materials.
• To demonstrate controlled co-packaging of subcompartments and functional macromolecules.
• To explore the potential for creating minimal synthetic cells capable of biochemical reactions.

Main Methods:

• Utilized bacteriophage P22 capsid as a host container.
• Co-encapsulated ferritin (Fn) cages and β-glycosidase enzyme (CelB) macromolecules.
• Employed controlled *in vitro* assembly to modulate stoichiometry and compartmentalization.

Main Results:

• Successfully constructed a nested protein cage system with co-packaged Fn and CelB within the P22 capsid.
• Demonstrated control over the degree of compartmentalization by adjusting Fn and CelB stoichiometry.
• Confirmed catalytic activity of the encapsulated CelB enzyme at high macromolecular concentrations.

Conclusions:

• Developed a novel approach for creating synthetic protein-based protocells.
• The system effectively confines functional proto-organelles and macromolecules, mimicking cellular environments.
• This model system holds potential for supporting a range of biochemical reactions within synthetic protocells.