2. Academic Validation
  3. Iso-ADP-Ribose Fluorescence Polarization Probe for the Screening of RNF146 WWE Domain Inhibitors

Iso-ADP-Ribose Fluorescence Polarization Probe for the Screening of RNF146 WWE Domain Inhibitors

  • ACS Chem Biol. 2024 Jan 18. doi: 10.1021/acschembio.3c00512.
Kewen Peng 1,  Ananya Anmangandla 1,  Sadhan Jana 1,  Yizhen Jin 2,  Hening Lin 3
Affiliations

Affiliations

  • 1 Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, United States.
  • 2 Graduate Program of Biochemistry, Molecular and Cell Biology, Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, United States.
  • 3 Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, United States.
PMID: 38237916 DOI: 10.1021/acschembio.3c00512
Abstract

Poly-ADP-ribosylation is an important protein post-translational modification with diverse biological consequences. After binding poly-ADP-ribose on axis inhibition protein 1 (AXIN1) through its WWE domain, RING finger protein 146 (RNF146) can ubiquitinate AXIN1 and promote its proteasomal degradation and thus the oncogenic Wnt signaling. Therefore, inhibiting the RNF146 WWE domain is a potential antitumor strategy. However, due to a lack of suitable screening methods, no inhibitors for this domain have been reported. Here, we developed a fluorescence polarization (FP)-based competition assay for the screening of RNF146 WWE inhibitors. This assay relies on a fluorescently tagged iso-ADP-ribose tracer compound, TAMRA-isoADPr. We report the design and synthesis of this tracer compound and show that it is a high-affinity tracer for the RNF146 WWE domain. This provides a convenient assay and will facilitate the development of small-molecule inhibitors for the RNF146 WWE domain.

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