A cell-permeable Ub-Dha probe for profiling E1-E2-E3 enzymes in live cells
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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:60 |
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| Enthalten in: |
Chemical communications (Cambridge, England) - 60(2024), 32 vom: 16. Apr., Seite 4342-4345 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Xia, Qiong [VerfasserIn] |
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| Links: |
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| Themen: |
EC 2.3.2.27 |
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| Anmerkungen: |
Date Completed 17.04.2024 Date Revised 17.04.2024 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1039/d4cc00415a |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a 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 | ||
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| 700 | 1 | |a Yuan, Rujing |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Pincheng |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Liwen |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Yi-Ming |e verfasserin |4 aut | |
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