Architecting Multicompartmentalized, Giant Vesicles with Recombinant Fusion Proteins.

Area of Science:

• Biotechnology
• Synthetic Biology
• Materials Science

Background:

• Designing artificial cells requires methods to create complex, compartmentalized structures.
• Existing methods often involve harsh chemicals or complex conjugation, limiting biomolecule encapsulation.

Purpose of the Study:

• To develop a straightforward strategy for constructing giant, multicompartmentalized vesicles using self-assembling recombinant fusion proteins.
• To demonstrate the encapsulation of diverse biomolecules and engineered compartments within these vesicles.
• To explore the potential of these vesicles as models for cellular processes and microreactor systems.

Main Methods:

• Utilized globule-zipper-elastin-like polypeptide fusion proteins for self-assembly in aqueous conditions.
• Employed the thin-film rehydration method for vesicle formation and encapsulation.
• Integrated water-soluble block copolymers to enhance vesicle stability and functionality.

Main Results:

• Successfully constructed giant, multicompartmentalized protein vesicles without organic solvents or chemical conjugation.
• Encapsulated various bioactive macromolecules, including proteins, coacervate droplets, and smaller vesicles.
• Demonstrated the ability of vesicles to support biochemically distinct reactions mediated by functional proteins.
• Enhanced structural stability and functional versatility through block copolymer integration.

Conclusions:

• The developed strategy offers a robust and versatile platform for creating cell-like structures using self-assembling proteins.
• These protein-assembled giant vesicles serve as advanced models for studying cellular organization and function.
• The findings highlight the potential of recombinant fusion proteins in advancing artificial cell design and microreactor technology.