Chemical Synthesis of Activity-Based Diubiquitin Probes

  • Guorui Li
  • Libo Yuan
  • Zhihao ZhuangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1513)


Activity-based diubiquitin probes are highly useful in probing the deubiquitinase (DUB) activity and ubiquitin chain linkage specificity. Here we describe a detailed protocol to synthesize a new class of diubiquitin DUB probes. In this method, two ubiquitin moieties are connected through a linker resembling the native linkage in size and containing a Michael acceptor for trapping the DUB active-site cysteine. Detailed procedures for generating the linker molecule are also described.

Key words

Activity-based probe Diubiquitin probe Chemical ligation DUB Ubiquitin chain Linkage specificity 



This work was supported, in part, by the US NIH grants GM097468 and NS085509 to Z. Zhuang.


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Chemistry and BiochemistryUniversity of DelawareNewarkUSA

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