Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity

Area of Science:

• Biochemistry
• Molecular Biology
• Optogenetics

Background:

• Allosteric regulation offers advantages for developing biological tools.
• Optogenetic applications of allosteric switches are limited.
• Spatiotemporal control of enzymatic activity is crucial for biological studies.

Purpose of the Study:

• To develop an optogenetic tool for precise spatiotemporal control of enzymatic activity.
• To engineer a Light-Regulated (LightR) allosteric switch module.
• To demonstrate the broad applicability of the LightR module across different enzymes.

Main Methods:

• Engineering of a Light-Regulated (LightR) allosteric switch module.
• Utilizing the tyrosine kinase Src as a model system.
• Modulating the LightR photoconversion cycle to tune off-kinetics.

Main Results:

• Demonstrated efficient light-mediated regulation of Src kinase activity.
• Identified temporally distinct signaling responses (seconds to minutes).
• Achieved light-controlled regulation of Abl, bRaf kinases, and Cre recombinase.

Conclusions:

• The engineered LightR module provides tight spatiotemporal control of enzymatic activity.
• LightR-Src off-kinetics are tunable, allowing for transient pulses and localized activity.
• The LightR module is broadly applicable for optogenetic control of diverse enzymes.