Spatiotemporal Control Over Protein Release from Artificial Cells via a Light-Activatable Protease

Area of Science:

• Biotechnology
• Synthetic Biology
• Chemical Biology

Background:

• Cellular protein regulation is crucial for intercellular communication and multicellularity.
• Mimicking this in artificial cells requires stimulus-responsive control over protein release.
• Existing artificial cell platforms lack precise regulatory mechanisms for protein secretion.

Purpose of the Study:

• To engineer a light-activatable system for spatiotemporal control of protein release from artificial cells.
• To develop a method for precise regulation of protein secretion in coacervate-based artificial cell platforms.
• To demonstrate light-activated protein transfer between artificial cell populations.

Main Methods:

• Development of a photocaged TEV protease (LaTEV) for light-activated protein release.
• Utilizing Ni2+-nitrilotriacetic acid moieties for sequestering His-tagged proteins within coacervates.
• Employing 365 nm light irradiation to activate LaTEV and cleave His-tags, releasing cargo proteins.
• Demonstrating selective and triggered protein release from coacervates via targeted light exposure.

Main Results:

• Successfully demonstrated photocaging and light-induced activation of LaTEV.
• Achieved precise spatiotemporal control over protein release rates from artificial cells.
• Showcased triggerable protein release from specific coacervates through selective irradiation.
• Validated light-activated directional protein transfer between distinct artificial cell populations.

Conclusions:

• The developed LaTEV system provides unprecedented light-based control over protein release in artificial cells.
• This platform enables engineering of light-responsive protein-mediated communication in synthetic systems.
• Advances the study of intercellular signaling and the development of novel protein delivery strategies.