Programmed spatial organization of biomacromolecules into discrete, coacervate-based protocells

Area of Science:

• Biotechnology
• Synthetic Biology
• Biochemistry

Background:

• Cell cytosol exhibits high biomacromolecular crowding, challenging to replicate in synthetic cells.
• Existing protocellular platforms have limitations in mimicking complex cellular environments.
• A need exists for biocompatible tools to accurately emulate the crowded cytosol.

Purpose of the Study:

• To develop a novel protocellular platform for controlled macromolecule loading.
• To create a tool that accurately mimics the crowded cellular environment.
• To enhance the functionality of synthetic cells.

Main Methods:

• Development of a discrete, membrane-bound coacervate-based protocellular platform.
• Utilized Ni2+-nitrilotriacetic acid and His-tagged proteins for controlled protein loading.
• Demonstrated protein accretion and functional enhancement of encapsulated systems.

Main Results:

• Achieved controlled, efficient, and benign loading of biologically relevant macromolecules.
• Enhanced an encapsulated two-enzyme cascade.
• Demonstrated protease-mediated cargo secretion.

Conclusions:

• The developed platform offers a versatile approach for programmed spatial organization of proteins.
• Expands the toolbox for protocell research and synthetic cell development.
• Paves the way for next-generation complex and regulated synthetic cells.