Assay systems for profiling deubiquitinating activity

Jinhong Cho, Jinyoung Park, Eunice Eunkyeong Kim, Eun Joo Song

Research output: Contribution to journalReview articlepeer-review

11 Scopus citations


Deubiquitinating enzymes regulate various cellular processes, particularly protein degradation, localization, and protein–protein interactions. The dysregulation of deubiquitinating enzyme (DUB) activity has been linked to several diseases; however, the function of many DUBs has not been identified. Therefore, the development of methods to assess DUB activity is important to identify novel DUBs, characterize DUB selectivity, and profile dynamic DUB substrates. Here, we review various methods of evaluating DUB activity using cell lysates or purified DUBs, as well as the types of probes used in these methods. In addition, we introduce some techniques that can deliver DUB probes into the cells and cell-permeable activity-based probes to directly visualize and quantify DUB activity in live cells. This review could contribute to the development of DUB inhibitors by providing important information on the characteristics and applications of various probes used to evaluate and detect DUB activity in vitro and in vivo.

Original languageEnglish
Article number5638
Pages (from-to)1-16
Number of pages16
JournalInternational Journal of Molecular Sciences
Issue number16
StatePublished - 2 Aug 2020

Bibliographical note

Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.


  • Activity-based probes
  • Cell permeability
  • Deubiquitination assay
  • Ub-probes


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