Browse by author
Lookup NU author(s): Professor Matthias TrostORCiD
Full text for this publication is not currently held within this repository. Alternative links are provided below where available.
© 2020, The Author(s), under exclusive licence to Springer Nature Limited.Deubiquitylating enzymes (DUBs) play a vital role in the ubiquitin pathway by editing or removing ubiquitin from their substrate. As breakthroughs within the ubiquitin field continue to highlight the potential of deubiquitylating enzymes as drug targets, there is increasing demand for versatile high-throughput (HT) tools for the identification of potent and selective DUB modulators. Here we present the HT adaptation of the previously published MALDI-TOF–based DUB assay method. In a MALDI-TOF DUB assay, we quantitate the amount of mono-ubiquitin generated by the in vitro cleavage of ubiquitin chains by DUBs. The method has been specifically developed for use with nanoliter-dispensing robotics to meet drug discovery requirements for the screening of large and diverse compound libraries. Contrary to the most common DUB screening technologies currently available, the MALDI-TOF DUB assay combines the use of physiological substrates with the sensitivity and reliability of the mass spectrometry–based readout.
Author(s): De Cesare V, Moran J, Traynor R, Knebel A, Ritorto MS, Trost M, McLauchlan H, Hastie CJ, Davies P
Publication type: Article
Publication status: Published
Journal: Nature Protocols
Year: 2020
Volume: 15
Pages: 4034–4057
Online publication date: 02/11/2020
Acceptance date: 26/08/2020
ISSN (print): 1754-2189
ISSN (electronic): 1750-2799
Publisher: Nature Research
URL: https://doi.org/10.1038/s41596-020-00405-0
DOI: 10.1038/s41596-020-00405-0
Altmetrics provided by Altmetric