생화학분자생물학회

	Name: Ji Min Lee ( jimin.lee@kaist.ac.kr )
	2022-present	Associate Professor, Graduate School of Medical Science & Engineering, KAIST
	2018-2022	Assistant Professor, Department of Molecular Biosciences, Kangwon National University
	2016-2018	Principal Scientist, Business Development Team, Samsung Bioepis
	2011-2015	Research Scientist, Therapeutic Antibody Group Samsung Advanced Institute of Technology
	2010-2011	Postdoctoral research fellow, Chromatin Dynamics Research Center, SNU
	2005-2010	Ph.D., Department of Biological Sciences, Seoul National University

Deubiquitinase Dynamics: Methodologies for Understanding Substrate Interactions

Deubiquitinases (DUBs) are essential regulators of protein homeostasis that influence cellular signaling, protein stability, and degradation by removing ubiquitin chains from substrate proteins. Understanding DUB–substrate interactions is critical to elucidate their functional roles and therapeutic potential. This review highlights key methodologies to investigate DUB activity and substrate interactions, including biochemical assays, fluorescence-based approaches, and in vitro deubiquitination assays. Biochemical methods, such as those measuring protein degradation rates, ubiquitination dynamics, and protein–protein interactions, provide valuable insights into DUB function and specificity. Fluorescence-based techniques that include photoconvertible reporters, fluorescent timers, and FRET enable the real-time monitoring of DUB dynamics and substrate turnover in live cells. Furthermore, in vitro deubiquitination assays provide direct mechanistic insights into DUB activity on target substrates. While each method provides unique insights, they also present challenges, like limited specificity or sensitivity, technical difficulties, or insufficient physiological relevance. Integrating complementary approaches can enhance accuracy and provide deeper insights into DUB–substrate interactions, facilitating the development of DUB-targeted therapeutic strategies.


BMB Rep. 2025 Mar 5. pii: 6444. [Epub ahead of print]
https://pubmed.ncbi.nlm.nih.gov/40058876/