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Updated: Sep 9, 2025

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
Synthesizing Defined Ubiquitin-Modified SUMO Dimers
Kai-Yu Hsu1, Yane-Shih Wang2,3
1Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
Abstract:
Ubiquitination and SUMOylation interactions between proteins play a critical role in various biological processes and the progression of diseases. To investigate the biological functions of the direct crosstalk between protein ubiquitination and SUMOylation biochemically, we have developed a method for synthesizing Ub-tagged SUMO2 dimers using an expanding genetic code approach. In this description, we outline the procedures for creating Ub-SUMO heterodimers through the incorporation of defined noncanonical amino acids (ncAAs) and biorthogonal functional group-guided conjugation techniques. These homogeneous Ub-SUMO heterodimers will serve as valuable tools for studying the mechanisms of Ub-SUMO chain elongation, degradation, topology, and other related protein-protein interactions.
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