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A pro-fluorescent ubiquitin-based probe to monitor cysteine-based E3 ligase activity

  • 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.
Metadaten
Verfasserangaben:David Aureliano Pérez BerrocalORCiD, Thimmalapura M. VishwanathaORCiD, Daniel Horn-GhetkoORCiD, J. Josephine BotschORCiD, Laura A. HehlORCiD, Sebastian KostrhonORCiD, Mohit MisraORCiDGND, Ivan ÐikićORCiDGND, Paul P. GeurinkORCiD, Hans van DamORCiD, Brenda A. SchulmanORCiDGND, Monique P. C. MulderORCiD
URN:urn:nbn:de:hebis:30:3-747162
DOI:https://doi.org/10.1002/anie.202303319
ISSN:1521-3773
Pubmed-Id:https://pubmed.ncbi.nlm.nih.gov/37272691
Titel des übergeordneten Werkes (Englisch):Angewandte Chemie / International ed.
Verlag:Wiley-VCH
Verlagsort:Weinheim
Dokumentart:Wissenschaftlicher Artikel
Sprache:Englisch
Datum der Veröffentlichung (online):05.06.2023
Datum der Erstveröffentlichung:05.06.2023
Veröffentlichende Institution:Universitätsbibliothek Johann Christian Senckenberg
Beteiligte Körperschaft:Gesellschaft Deutscher Chemiker
Datum der Freischaltung:18.07.2023
Freies Schlagwort / Tag:Biological Activity; Drug Discovery; E3-Ligases; Fluorescent Probes; Profiling Transthiolation Activity
Jahrgang:2023
Ausgabe / Heft:online version before inclusion in an issue
Aufsatznummer:e202303319
Seitenzahl:12
Bemerkung:
Early View: Online Version before inclusion in an issue
Bemerkung:
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.
HeBIS-PPN:511283903
Institute:Medizin / Medizin
Fachübergreifende Einrichtungen / Buchmann Institut für Molekulare Lebenswissenschaften (BMLS)
DDC-Klassifikation:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Lizenz (Deutsch):License LogoCreative Commons - CC BY - Namensnennung 4.0 International