In situ assembly of macromolecular complexes triggered by light

Area of Science:

• Chemical Biology
• Molecular Engineering
• Biophysics

Background:

• Precise control over protein complex organization in time and space is crucial for understanding cellular processes.
• Existing methods for protein manipulation often lack spatiotemporal resolution or site-specificity.

Purpose of the Study:

• To develop a novel light-triggered molecular recognition system for controlled protein complex organization.
• To achieve site-specific, reversible, and high-affinity binding to His-tagged proteins using external light stimuli.

Main Methods:

• Development of a self-inactivated, lock-and-key recognition element.
• Photocleavage of an intramolecular ligand to activate a multivalent chelator head.
• Demonstration of binding in solution and at interfaces using laser scanning microscopy.
• Iterative in situ writing and binding for multiplexed organization.

Main Results:

• Achieved light-triggered binding to His-tagged proteins with tunable affinity (zero to nanomolar).
• Demonstrated site-specific, stable, and reversible binding with high temporal and spatial resolution.
• Successfully organized protein complexes in a multiplexed manner using an iterative in situ process.

Conclusions:

• The developed light-triggered molecular recognition system enables precise spatiotemporal control over protein-protein interactions.
• This technology facilitates light-induced protein clustering, offering a powerful tool for manipulating cellular processes.
• Potential applications in chemical biology for dynamic control of molecular assemblies and cellular functions.