The SpyTag-SpyCatcher protein conjugation system has recently gained popularity due to its fast kinetics and high yield under biologically favorable conditions in both in vitro and in vivo settings. We can further expand the utility of this system by introducing the ability to spatially and temporally control the conjugation event. For intracellular applications, the light-oxygen-voltage 2 domain of Avena sativa (AsLOV2) has been used for modulating the affinity of protein-peptide ligand interactions in response to blue light. We have thus genetically fused the SpyTag to the AsLOV2 domain to create a Blue Light Inducible SpyCatcher System (BLISS). In the dark state, the SpyTag is blocked from reacting with the SpyCatcher, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag. We have demonstrated the ability to use light to biologically pattern surfaces with proteins. We have also applied this system for the optogenetic rescue of intracellular proteins, where a protein targeted towards degradation is cleaved from its degron tag in response to the BLISS protein conjugation event. We anticipate the precise control our strategy provides will be a strong tool for making protein microarrays and crafting biomaterial composition as well as controlling therapeutic protein activity in select cells.

Engineering a blue light inducible SpyCatcher system (BLISS) for in vitro photo-pattering of proteins and optically controlled protein rescue

255th National Meeting (2018)

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