Single-Protein-Specific Redox Targeting in Live Mammalian Cells and C. elegans.

ABSTRACT: T-REX (targetable reactive electrophiles and oxidants) enables electrophile targeting in living systems with high spatiotemporal precision and at single-protein-target resolution. T-REX allows functional consequences of individual electrophile signaling events to be directly linked to on-target modifications. T-REX is accomplished by expressing a HaloTagged protein of interest (POI) and introducing a Halo-targetable bioinert photocaged precursor to a reactive electrophilic signal (RES). Light exposure releases the unfettered RES on demand, enabling precision modification of the POI due to proximity. Using alkyne-functionalized 4-hydroxynonenal (HNE) as a representative RES, this protocol delineates optimized strategies to (1) execute T-REX in live human cells and C. elegans, (2) quantitate

SUBMITTER: Van Hall-Beauvais A 

PROVIDER: S-EPMC6125161 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature