A Double Photocage Strategy to Construct Light-Controllable and Spatiotemporally Trackable Cathepsin B Activity-Based Probes

Utilizing multiple cages to selectively modulate the activity of biomolecules is indispensable to achieving controllable and trackable activity manipulation. However, trackable cages that can be used to monitor the activation of biomolecules are rare. In this work, we utilized a double photocage strategy to achieve light-controllable and spatiotemporally trackable activation. To demonstrate biological applicability, we used the well-known cancer cell biomarker cathepsin B as the target and constructed double photocaged cathepsin B activity-based probe 2PPG-FK-AcRha that performed well in cancer cell cultures. Using our probe, we could monitor the light-activation by the blue fluorescence of 7-diethylamino-4-hydroxymethyl-coumarin (DEACM) and simultaneously probe the activity of cathepsin B through the green fluorescence of acetyl rhodamine (AcRha). Additionally, by partially irradiating the cell cultures, the regional photoactivation experiments also demonstrated great spatial controllability and trackability of our probe.