Rapid Covalent-Probe Discovery by Electrophile-Fragment Screening

Efrat Resnick, Anthony Bradley, Jinrui Gan, Alice Douangamath, Tobias Krojer, Ritika Sethi, Paul P. Geurink, Anthony Aimon, Dom Bellini, James Bennett, Michael Fairhead, Oleg Fedorov, Ronen Gabizon, Jin Gan, Jingxu Gu, Nava Reznik, Gian Filippo Ruda, Laura Diaz-Saez, Verena M. Straub, Tamas SzommerSrikannathasan Velupillai, Daniel Zaidman, Yanling Zhang, Alun R. Coker, Christopher G. Dowson, Chu Wang, Kilian V. M. Huber, Paul E. Brennan, Huib Ovaa, Frank von Delft, Nir London

Research output: Contribution to journalArticlepeer-review


Covalent probes can display unmatched potency, selectivity, and duration of action; however, their discovery is challenging. In principle, fragments that can irreversibly bind their target can overcome the low affinity that limits reversible fragment screening, but such electrophilic fragments were considered nonselective and were rarely screened. We hypothesized that mild electrophiles might overcome the selectivity challenge and constructed a library of 993 mildly electrophilic fragments. We characterized this library by a new high-throughput thiol-reactivity assay and screened them against 10 cysteine-containing proteins. Highly reactive and promiscuous fragments were rare and could be easily eliminated. In contrast, we found hits for most targets. Combining our approach with high-throughput crystallography allowed rapid progression to potent and selective probes for two enzymes, the deubiquitinase OTUB2 and the pyrophosphatase NUDT7. No inhibitors were previously known for either. This study highlights the potential of electrophile-fragment screening as a practical and efficient tool for covalent-ligand discovery.

Original languageEnglish
Pages (from-to)8951-8968
Number of pages18
JournalJournal of the American Chemical Society
Issue number22
StatePublished - 5 Jun 2019

All Science Journal Classification (ASJC) codes

  • General Chemistry
  • Biochemistry
  • Catalysis
  • Colloid and Surface Chemistry


Dive into the research topics of 'Rapid Covalent-Probe Discovery by Electrophile-Fragment Screening'. Together they form a unique fingerprint.

Cite this