Neutron-encoded diubiquitins to profile linkage selectivity of deubiquitinating enzymes
- Author(s)
- van Tol, BDM; van Doodewaerd, BR; Lageveen-Kammeijer, GSM; Jansen, BC; Talavera Ormeño, CMP; Hekking, PJM; Sapmaz, A; Kim, RQ; Moutsiopoulou, A; Komander, D; Wuhrer, M; van der Heden van Noort, GJ; Ovaa, H; Geurink, PP;
- Details
- Publication Year 2023-03-25,Volume 14,Issue #1,Page 1661
- Journal Title
- Nature Communications
- Abstract
- Deubiquitinating enzymes are key regulators in the ubiquitin system and an emerging class of drug targets. These proteases disassemble polyubiquitin chains and many deubiquitinases show selectivity for specific polyubiquitin linkages. However, most biochemical insights originate from studies of single diubiquitin linkages in isolation, whereas in cells all linkages coexist. To better mimick this diubiquitin substrate competition, we develop a multiplexed mass spectrometry-based deubiquitinase assay that can probe all ubiquitin linkage types simultaneously to quantify deubiquitinase activity in the presence of all potential diubiquitin substrates. For this, all eight native diubiquitins are generated and each linkage type is designed with a distinct molecular weight by incorporating neutron-encoded amino acids. Overall, 22 deubiquitinases are profiled, providing a three-dimensional overview of deubiquitinase linkage selectivity over time and enzyme concentration.
- Publisher
- NPG
- Research Division(s)
- Ubiquitin Signalling
- PubMed ID
- 36966155
- Publisher's Version
- https://doi.org/10.1038/s41467-023-37363-6
- Open Access at Publisher's Site
https://doi.org/10.1038/s41467-023-37363-6- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2023-04-13 11:09:56
Last Modified: 2023-04-13 11:57:05