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Engineering a Blue Light Inducible SpyTag System (BLISS).

Emily J Hartzell¹, Justin Terr¹, Wilfred Chen¹

¹Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware 19716, United States.

Journal of the American Chemical Society

|June 2, 2021

Summary

Researchers developed a blue light-inducible protein conjugation system (BLISS) using SpyTag and light-oxygen-voltage domain 2 (AsLOV2). This system allows for precise spatial and temporal control over protein reactions, enhancing biofabrication and optogenetics.

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Area of Science:

Biochemistry
Molecular Biology
Optogenetics

Background:

The SpyCatcher/SpyTag system is a widely used protein conjugation method known for its efficiency.
Controlling the timing and location of protein conjugation is crucial for advanced biological applications.
Natural photoreceptor proteins offer mechanisms for light-dependent biological control.

Purpose of the Study:

To engineer a light-inducible protein conjugation system for enhanced spatial and temporal control.
To integrate the light-oxygen-voltage domain 2 (AsLOV2) of Avena sativa into the SpyTag system.
To create a blue light-inducible SpyTag system (BLISS) for controllable protein conjugation.

Main Methods:

Engineered variants of SpyTag by inserting it into different locations of the AsLOV2 Jα-helix.
Characterized the kinetics and dynamic range of the BLISS variants under varying light conditions.
Demonstrated spatial control using photopatterning of fluorescent proteins.

Main Results:

Developed BLISS variants with dynamic ranges exceeding 15, enabling tunable protein conjugation.
Showcased that conjugation is blocked in the dark and rapidly activated by blue light.
Confirmed instantaneous quenching of the reaction upon light removal.
Successfully demonstrated spatial control via photopatterning.

Conclusions:

The BLISS system provides a novel method for light-dependent protein conjugation.
This system offers precise spatial and temporal control over biochemical reactions.
BLISS has significant potential for applications in biofabrication, optogenetics, and synthetic biology.