Strategies for Monitoring “Ubiquitin C-Terminal Hydrolase 1” (Yuh1) Activity

Shahaf Saad, Eden Berda, Yuval Klein, Suha Issa, Elah Pick

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review


The family of ubiquitin C-terminal hydrolases (UCHs(releases ε-linked amide bonds positioned at the C-terminus of ubiquitin. UCHL3 is a highly conserved and dual functional member of this family, recognizing C-terminal extensions of two paralogous modifiers: ubiquitin and NEDD8. The Saccharomyces cerevisiae orthologue of UCHL3, namely, Yuh1, is the only UCH family member in this organism. Like UCHL3, Yuh1 recognizes ubiquitin as well as Rub1, the direct orthologue of NEDD8 in S. cerevisiae. We describe here a method for examining the activity of bacteria and yeast expressed Yuh1 by monitoring the C-terminal trimming of UBB + 1 and Rub1 + 1 through immunoblotting and the increased AMC fluorescence readout detected through a plate reader.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages16
StatePublished - 2023

Publication series

NameMethods in molecular biology (Clifton, N.J.)
ISSN (Print)1064-3745

Bibliographical note

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.


  • Amides
  • Proteolysis
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae/genetics
  • Ubiquitin
  • Ubiquitin Thiolesterase
  • Ubiquitins


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