An activity-dependent proximity ligation platform for spatially resolved quantification of active enzymes in single cells.

ABSTRACT: Integration of chemical probes into proteomic workflows enables the interrogation of protein activity, rather than abundance. Current methods limit the biological contexts that can be addressed due to sample homogenization, signal-averaging, and bias toward abundant proteins. Here we report a platform that integrates family-wide chemical probes with proximity-dependent oligonucleotide amplification and imaging to quantify enzyme activity in native contexts with high spatial resolution. Application of this method, activity-dependent proximity ligation (ADPL), to serine hydrolase and cysteine protease enzymes enables quantification of differential enzyme activity resulting from endogenous changes in localization and expression. In a competitive format, small-molecule target engagement with e

SUBMITTER: Li G 

PROVIDER: S-EPMC5701173 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature