PFI-7 is a potent, selective and cell-active chemical probe inhibitor for Glucose-Induced Degradation 4 (GID4), the substrate receptor of the CTLH ubiquitin ligase complex, antagonizes Pro/N-degron binding to human GID4 with SPR Kd of 79 nM.
PFI-7 can directly displace the Pro/N-degron peptide PGLWKS from the GID4-binding pocket in a peptide displacement assay (Kdisp=4.1 uM).
PFI-7 binds within the β-barrel of GID4 substrate binding pocket, disrupting its interaction with the canonical Pro/N-degron peptide in cells.
PFI-7 decreases GID4-HaloTag binding to PGLKWS-NanoLuc on human embryonic kidney 293T (HEK293T) cells with IC50 of 0.54 uM.
PFI-7 is a suitable tool compound for use in cellular studies to interrogate GID4-mediated recruitment to the CTLH complex.
The Pro/N-degron pathway is a recently identified pathway that recognizes N-terminal degrons containing unmodified Proline residues (Pro/N-degrons).