A cell-permeable Ub-Dha probe for profiling E1-E2-E3 enzymes in live cells.
Qiong XiaXianbin MengYu WangRujing YuanPincheng LiLiwen LiuYi-Ming LiPublished in: Chemical communications (Cambridge, England) (2024)
Activity-based ubiquitin probes (Ub-ABPs) have recently been developed as effective tools for studying the capabilities of E1-E2-E3 enzymes, but most of them can only be used in cell lysates. Here, we report the first cell-penetrating Ub-Dha probes based on thiazolidine-protected cysteines, which enable successful delivery into cells confirmed by a fluorophore at the N-terminus of Ub and live-cell fluorescence microscopy. A total of 18 E1-E2-E3 enzymes in live cells were labelled and enriched in combination with label-free quantification (LFQ) mass spectrometry. This work provided a new cell-penetrating Ub tool for studying the activity and function of Ub-related enzymes.
Keyphrases
- single cell
- induced apoptosis
- mass spectrometry
- cell therapy
- cell cycle arrest
- small molecule
- label free
- single molecule
- high resolution
- living cells
- stem cells
- oxidative stress
- signaling pathway
- endoplasmic reticulum stress
- mesenchymal stem cells
- quantum dots
- fluorescence imaging
- high performance liquid chromatography
- pi k akt