biostudies-literatureJones KAYhim FoundationNIMH NIH HHSNational Institute of Mental HealthNational Institute of General Medical SciencesNIGMS NIH HHS1768-1776https://www.ebi.ac.uk/biostudies/studies/S-EPMC6891849biostudies-literatureUnknown5(11)Split reporters based on fluorescent proteins and luciferases have emerged as valuable tools for measuring interactions in biological systems. Relatedly, biosensors that transduce measured input signals into outputs that influence the host system are key components of engineered gene circuits for synthetic biology applications. While small-molecule-based imaging agents are widely used in biological studies, and small-molecule-based drugs and chemical probes can target a range of biological processes, a general method for generating a target small molecule in a biological system based on a measured input signal is lacking. Here, we develop a proximity-dependent split esterase that selectively unmasks ester-protected small molecules in an interaction-dependent manner. Exploiting the versatilACS central scienceDevelopment of a Split Esterase for Protein-Protein Interaction-Dependent Small-Molecule Activation.PMC6891849RF1 MH114102T32 GM008720R35 GM119840Dickinson BCLippert ARBeck MWLewis JCAn WJones KAKentala KfalseDevelopment of a Split Esterase for Protein-Protein Interaction-Dependent Small-Molecule Activation.Split reporters based on fluorescent proteins and luciferases have emerged as valuable tools for measuring interactions in biological systems. Relatedly, biosensors that transduce measured input signals into outputs that influence the host system are key components of engineered gene circuits for synthetic biology applications. While small-molecule-based imaging agents are widely used in biological studies, and small-molecule-based drugs and chemical probes can target a range of biological processes, a general method for generating a target small molecule in a biological system based on a measured input signal is lacking. Here, we develop a proximity-dependent split esterase that selectively unmasks ester-protected small molecules in an interaction-dependent manner. Exploiting the versatil2019-01-01T00:00:00Z2019 Nov2026-05-02T05:49:10.793Z2020-05-21T22:37:48ZS-EPMC68918493180767810.1021/acscentsci.9b00567