Genetically encoded photocatalytic protein labeling enables spatially-resolved profiling of intracellular proteome.

ABSTRACT: Mapping the subcellular organization of proteins is crucial for understanding their biological functions. Herein, we report a reactive oxygen species induced protein labeling and identification (RinID) method for profiling subcellular proteome in the context of living cells. Our method capitalizes on a genetically encoded photocatalyst, miniSOG, to locally generate singlet oxygen that reacts with proximal proteins. Labeled proteins are conjugated in situ with an exogenously supplied nucleophilic probe, which serves as a functional handle for subsequent affinity enrichment and mass spectrometry-based protein identification. From a panel of nucleophilic compounds, we identify biotin-conjugated aniline and propargyl amine as highly reactive probes. As a demonstration of the spatial specificit

SUBMITTER: Zheng F 

PROVIDER: S-EPMC10205723 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature