Dynamic Light-Induced Protein Patterns at Model Membranes.

Area of Science:

• Synthetic Biology
• Biochemistry
• Cell Biology

Background:

• Precise protein localization and activation on cell membranes are crucial for cellular processes like polarization, migration, and division.
• Controlling these processes in synthetic cells requires methods for protein recruitment with high spatial and temporal resolution.

Purpose of the Study:

• To develop a method for fabricating light-regulated, reversible protein patterns on lipid membranes with high spatiotemporal precision.
• To enable dynamic control over protein localization for applications in synthetic cell biology.

Main Methods:

• Immobilization of the photoswitchable protein iLID (improved light-inducible dimer) on supported lipid bilayers (SLBs) and giant unilamellar vesicles (GUVs).
• Utilizing blue light illumination to trigger iLID binding to its partner Nano (wild-type SspB).
• Recruiting proteins of interest (POIs) fused to Nano from solution to illuminated membrane areas.

Main Results:

• Demonstrated successful immobilization of iLID on both SLBs and GUVs.
• Achieved light-induced, spatially precise recruitment of Nano-fused proteins to illuminated membrane regions.
• Confirmed the reversibility of protein binding in the dark, allowing dynamic release.

Conclusions:

• The developed method offers a flexible and versatile approach for precisely controlling protein localization in space and time using blue light.
• This technique is valuable for engineering synthetic cells and studying membrane-associated cellular processes.