Photocleavable dimerizer for the rapid reversal of molecular trap antagonists (597.3)

Abstract

Developing a molecular tool that can rapidly turn on and off an acute antagonist will be immensely helpful to understand the initiation of a cellular program. Herein, we report the development of a photocleavable analog of AP20187, a cell permeable molecule used to dimerize FKBP fusion proteins and initiate biological signaling cascades, gene expression or disrupt protein‐protein interactions. We demonstrate that this reagent in conjugation with a molecular trap permits the ability to rapidly and specifically antagonize a molecular interaction in vitro and follow a biological process due to this acute antagonism (e.g., endosome dispersion). More importantly, photocleavage of the analog permits the unique ability to follow the cell’s return to homeostasis. Herein, we demonstrated the fine temporal control of endosome dispersion and restoration using an LC8 trap. We posit that this photocleavable AP20187 analog can be used in other systems where the dimerization of FKBP has been used to initiate signaling pathways (e.g., kinases). In toto, the combination of a molecular trap and the photocleavable analog opens up new avenues to study signaling events within a single cell in a whole organism.Grant Funding Source: Supported by the generous support of R21NS071166 and also by P30 CA033572 from the NCI