Issue 32, 2024
From the journal:

Chemical Communications

A cell-permeable Ub-Dha probe for profiling E1–E2–E3 enzymes in live cells†

Qiong Xia,‡a   Xianbin Meng,‡b   Yu Wang,*a   Rujing Yuan,a   Pincheng Li,a   Liwen Liua  and  Yi-Ming Li ORCID logo *a  

* Corresponding authors

a School of Food and Biological Engineering, Engineering Research Center of Bio-process, Ministry of Education, Hefei University of Technology, Hefei 230009, China
E-mail: ymli@hfut.edu.cn, wangyuch@mail.ustc.edu.cn

b MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China

Abstract

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.

Graphical abstract: A cell-permeable Ub-Dha probe for profiling E1–E2–E3 enzymes in live cells

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Article information

DOI
https://doi.org/10.1039/D4CC00415A
Article type
Communication
Submitted
30 Jan 2024
Accepted
19 Mar 2024
First published
21 Mar 2024

Chem. Commun., 2024,60, 4342-4345

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A cell-permeable Ub-Dha probe for profiling E1–E2–E3 enzymes in live cells

Q. Xia, X. Meng, Y. Wang, R. Yuan, P. Li, L. Liu and Y. Li, Chem. Commun., 2024, 60, 4342 DOI: 10.1039/D4CC00415A

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