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Updated: Jan 24, 2026

In Vitro Analysis of E3 Ubiquitin Ligase Function
Published on: May 14, 2021
An activity-based probe developed by a sequential dehydroalanine formation strategy targets HECT E3 ubiquitin ligases
1Department of Chemistry, University of Science and Technology of China, Hefei 230026, China. shijing@ustc.edu.cn.
Abstract:
E3 ligases play a critical role in ubiquitin (Ub) conjugation cascades, and any aberration in their activity is associated with a number of diseases. Advancement in our knowledge of understanding the roles of HECT E3s requires biochemical tools such as activity-based probes (ABPs). In this study we developed a novel dehydroalanine (Dha)-based E2-Ub ABP using a strategy that is a combination of practical hydrazide-based native chemical ligation and sequential Dha formation. The probe could be used for labeling HECT E3s not only in vitro but also in endogenous cellular contexts. Our easy-to-implement method is expected to be useful for the preparation of Dha based Ub family E2 conjugate ABPs.
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