Chemo-optogenetic Protein Translocation System Using a Photoactivatable Self-Localizing Ligand.

Area of Science:

• Cell Biology
• Biochemistry
• Optogenetics

Background:

• Light-based manipulation of subcellular protein localization offers precise spatiotemporal control of cellular signaling.
• Current methods often rely on light-induced protein heterodimerization, limiting flexibility.
• A need exists for strategies using small molecules to control protein localization upon light activation without requiring additional proteins.

Purpose of the Study:

• To develop and validate a chemo-optogenetic approach for light-induced protein translocation using a synthetic molecule.
• To demonstrate the ability of this system to control protein localization and cellular signaling with high precision.
• To establish a versatile platform for interrogating and engineering dynamic cellular functions.

Main Methods:

• Development of a photoactivatable self-localizing ligand (paSL).
• Utilizing paSL to recruit tagged proteins from the cytoplasm to the plasma membrane upon light illumination.
• Characterization of the paSL-induced protein translocation (paSLIPT) system's reversibility and spatiotemporal control in living cells.

Main Results:

• Successfully engineered a paSL capable of rapid, light-induced recruitment of tagged proteins to the plasma membrane.
• Demonstrated reversible and spatiotemporally controlled protein translocation (paSLIPT) in living cells.
• Showcased the system's utility for simultaneous optical stimulation and multiplexed imaging within single cells.

Conclusions:

• The developed paSLIPT system provides a novel and effective chemo-optogenetic platform for precise control of protein localization.
• This approach enables sophisticated interrogation and engineering of dynamic cellular processes with high spatiotemporal resolution.
• paSLIPT offers advantages over existing methods, paving the way for new applications in cell biology and synthetic biology.