TY  - JOUR
A1  - Pérez Berrocal, David Aureliano
A1  - Vishwanatha, Thimmalapura M.
A1  - Horn-Ghetko, Daniel
A1  - Botsch, J. Josephine
A1  - Hehl, Laura A.
A1  - Kostrhon, Sebastian
A1  - Misra, Mohit
A1  - Ðikić, Ivan
A1  - Geurink, Paul P.
A1  - Dam, Hans van
A1  - Schulman, Brenda A.
A1  - Mulder, Monique P. C.
T1  - A pro-fluorescent ubiquitin-based probe to monitor cysteine-based E3 ligase activity
T2  - Angewandte Chemie / International ed.
N2  - Protein post-translational modification with ubiquitin (Ub) is a versatile signal regulating almost all aspects of cell biology, and an increasing range of diseases is associated with impaired Ub modification. In this light, the Ub system offers an attractive, yet underexplored route to the development of novel targeted treatments. A promising strategy for small molecule intervention is posed by the final components of the enzymatic ubiquitination cascade, E3 ligases, as they determine the specificity of the protein ubiquitination pathway. Here, we present UbSRhodol, an autoimmolative Ub-based probe, which upon E3 processing liberates the pro-fluorescent dye, amenable to profile the E3 transthiolation activity for recombinant and in cell-extract E3 ligases. UbSRhodol enabled detection of changes in transthiolation efficacy evoked by enzyme key point mutations or conformational changes, and offers an excellent assay reagent amenable to a high-throughput screening setup allowing the identification of small molecules modulating E3 activity.
KW  - Biological Activity
KW  - Drug Discovery
KW  - Fluorescent Probes
KW  - E3-Ligases
KW  - Profiling Transthiolation Activity
Y1  - 2023
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/74716
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-747162
SN  - 1521-3773
N1  - Early View: Online Version before inclusion in an issue
N1  - This work was supported by the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 765445 to D.P.B., the EU/EFPIA/OICR/McGill/KTH/Diamond Innovative Medicines Initiative 2 Joint Undertaking (EUbOPEN grant no. 875510) and NWO (VIDI Grant VI. 213.110 to M.P.C.M.). Work in B.A.S.’s lab was funded by the ERC under the H2020 research and innovation programme (ADG-789016-Nedd8Activate), by the DFG (SCHU3196/1-1, Gottfried Wilhelm Leibniz Prize), and the Max Planck Society.
VL  - 2023
IS  - online version before inclusion in an issue
PB  - Wiley-VCH
CY  - Weinheim
ER  -