A selective, turn-on fluorescent probe for live-cell super-resolution imaging of c-Src kinase

23 May 2024, Version 1
This content is a preprint and has not undergone peer review at the time of posting.


Subcellular localization is important in regulating kinase activation and function. Small molecule flu-orescent probes for kinases can provide compli-mentary advantages to immunofluorescence and genetically encoded tags. Here, we developed a highly selective fluorescent probe for c-Src, a non-receptor tyrosine kinase. Our probe combines a py-razolopyrimidine (PP) kinase binding group and a coumarin fluorophore, and covalently binds to c-Src through a non-conserved cysteine (Cys-280). Our PP-coumarin (PP-C) probe is a turn-on fluo-rescent probe of c-Src that can be used in live-cell microscopy without washing and is compatible with super-resolution live-cell STED microscopy. We used PP-C to profile c-Src localization and expres-sion level in a panel of triple-negative breast cancer (TNBC) cell lines.


live cell imaging

Supplementary materials

Supplementary Information
Materials and methods, synthesis of PP-C, supplementary figures (PDF).


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