摘要
Spatiotemporal profiling of nuclear-associated proteomes is crucial for elucidating disease mechanisms,identifying key therapeutic targets,and guiding the design of effective drugs.Currently,proximity labeling(PL)using genetically transfected enzymes or photocatalyst-based probes has emerged as a powerful tool for proteomic mapping.However,these approaches are limited by their incompatibility with hard-to-transfect cells and primary tissues,as well as by the lack of efficient nucleus-targeting strategies.In this study,we developed a photocatalytic PL strategy(Pc-PL)that enables efficient enrichment of nuclear-associated proteins by combining a nucleus-targeted photosensitizer(NCP)with photocatalysis-mediated reactive biotin labeling.Compared with traditional photocatalysts such as chlorin e6 and rose Bengal,NCP exhibited superior nuclear accumulation across various cell types.Cellular experiments confirmed that NCP-mediated photoactivation precisely localized biotin labeling within the nucleus,enabling selective enrichment of nuclear proteins via subsequent streptavidin-based magnetic capture.Coupling Pc-PL with quantitative mass spectrometry enabled highresolution mapping of nuclear proteomes and led to the discovery of previously unrecognized senescence-associated regulators,including TMPO.Collectively,these findings establish Pc-PL as an innovative and versatile tool for highresolution nuclear proteomics,offering broad potential for target discovery and drug development.
基金
financial support of the National Natural Science Foundation of China(32501182)
the Guangdong Provincial Department of Education Natural Science Research Project(2025ZDZX4030).
