A Robust, GFP-Orthogonal Photoswitchable Inhibitor Scaffold Extends Optical Control over the Microtubule Cytoskeleton

•SBTub3 photocontrols microtubule dynamics, organization, and dependent processes •Microtubule photocontrol is cell sub-cellularly precise temporally reversible •SBT orthogonal to GFP/YFP imaging SBTs are metabolically stable •The SBT scaffold promising for photopharmaceuticals other protein targets Optically controlled chemical reagents, termed “photopharmaceuticals,” powerful tools spatiotemporal control of proteins particularly when genetic methods, such as knockouts or optogenetics not viable options. However, current photopharmaceutical scaffolds, azobenzenes intolerant labile, posing severe limitations biological use. We rationally designed a photoswitchable "SBT" overcome these problems, then derivatized it create exceptionally robust fully GFP/YFP-orthogonal "SBTub" tubulin inhibitors. Lead compound SBTub3 allows reversible, cell-precise, even subcellularly photomodulation microtubule-dependent processes. By overcoming the previous photopharmaceuticals, SBTubs offer applications in biology, their robustness druglikeness favorable intracellular vivo applications. furthermore expect that orthogonality will inspire derivatizations directed at extending range targets.