Modulating Protein Function through Genetically Encoded Oxidative Chemistry.

Oxidative chemistry underlies many endogenous signaling pathways but remains underutilized as a programmable strategy for regulating protein function in living cells. Here, we establish genetically encoded oxidative chemistry as a tunable framework for modulating diverse proteins by coupling a photosensitizer to defined intracellular contexts. Using miniSOG to generate reactive oxygen species (ROS), we show that controlled intracellular oxidation increases the fluorescence of the redox reporter HyPerRed and activates redox-sensitive TRP ion channels, with strong responses in TRPA1 and TRPV1 but not TRPV4. Pathway-selective scavengers reveal differential coupling of soluble and membrane targets to distinct oxidative processes, supporting selectivity by context rather than uniform oxidative perturbation. Modulation strength and kinetics are quantitatively tunable through illumination parameters, expression ratios, and subcellular localization, with membrane targeting enhancing the coupling to membrane effectors. Finally, fusion targeting of miniSOG to TRPV1 and modulation of endogenous TRPA1 in human fibroblasts extend this approach to protein-proximal and native cellular settings. Together, these results position genetically encoded oxidative chemistry as a versatile and spatially organized modality for engineering protein function in living cells within a defined operating regime.