Three-Dimensional Protein Networks Assembled by Two-Photon Activation

Spatial and temporal control over chemical and biological processes plays a key role in life and material sciences. Here we synthesized a two‐photon‐activatable glutathione (GSH) to trigger the interaction with glutathione S‐transferase (GST) by light at superior spatiotemporal resolution. The compound shows fast and well‐confined photoconversion into the bioactive GSH, which is free to interact with GST‐tagged proteins. The GSH/GST interaction can be phototriggered, changing its affinity over several orders of magnitude into the nanomolar range. Multiplexed three‐dimensional (3D) protein networks are simultaneously generated in situ through two‐photon fs‐pulsed laser‐scanning excitation. The two‐photon activation facilitates the three‐dimensional assembly of protein structures in real time at hitherto unseen resolution in time and space, thus opening up new applications far beyond the presented examples. Chemical biology aims at controlling protein interactions and cell behavior by external stimuli. Tools are needed to manipulate and structure target molecules in a non‐invasive manner, and this is best achieved by light. A two‐photon‐activatable glutathione can be used for the light‐guided three‐dimensional patterning of glutathione S‐transferase at superior spatiotemporal resolution.