Photocrosslinking activity-based probes for ubiquitin RING E3 ligases

Mathur, S., Fletcher, A. J., Branigan, E., Hay, R. T. and Virdee, S. (2019) Photocrosslinking activity-based probes for ubiquitin RING E3 ligases. Cell Chemical Biology, 27(1), 74-82.e6. (doi: 10.1016/j.chembiol.2019.11.013) (PMID:31859248) (PMCID:PMC6963778)

Abstract

Summary: Activity-based protein profiling is an invaluable technique for studying enzyme biology and facilitating the development of therapeutics. Ubiquitin E3 ligases (E3s) are one of the largest enzyme families and regulate a host of (patho)physiological processes. The largest subtype are the RING E3s of which there are >600 members. RING E3s have adaptor-like activity that can be subject to diverse regulatory mechanisms and have become attractive drug targets. Activity-based probes (ABPs) for measuring RING E3 activity do not exist. Here we re-engineer ubiquitin-charged E2 conjugating enzymes to produce photocrosslinking ABPs. We demonstrate activity-dependent profiling of two divergent cancer-associated RING E3s, RNF4 and c-Cbl, in response to their native activation signals. We also demonstrate profiling of endogenous RING E3 ligase activation in response to epidermal growth factor (EGF) stimulation. These photocrosslinking ABPs should advance E3 ligase research and the development of selective modulators against this important class of enzymes.

• Item Type: Articles
• Additional Information: This work was funded by the Biotechnology and Biological Sciences Research Council ( BB/P003982/1 ); UK Medical Research Council ( MC_UU_12016/8 ); Wellcome Trust , United Kingdom, Senior Investigator award (098391/Z/12/Z) to R.T.H
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Fletcher, Dr Adam
• Authors: Mathur, S., Fletcher, A. J., Branigan, E., Hay, R. T., and Virdee, S.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Cell Chemical Biology
• Publisher: Elsevier (Cell Press)
• ISSN: 2451-9456
• ISSN (Online): 2451-9448
• Copyright Holders: Copyright © 2019 The Authors
• First Published: First published in Cell Chemical Biology 27:(1):74-82.e6
• Publisher Policy: Reproduced under a Creative Commons Licence