| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | A gene encoding full-length KLHDC2 (Uniprot: Q9Y2U9) with an N-terminal GST-TEV-His tag was synthesized (Genscript) and cloned into a pFastBac vector./. Full-length EloB (Uniprot: Q15370) and EloC (Uniprot: Q15369) were synthesized (Genscript) and cloned into multiple cloning sites of pFastBacDual. /The final reactions included 3 nM 6×His-KLHDC2KD, 2 nM biotinylated SelK peptide (biotin-HLRGSPPPMAGG, Genscript), 12.5 µg mL–1 Anti-6×His Acceptor Beads (PerkinElmer), and 12.5 µg mL–1 Streptavidin Donor Beads (PerkinElmer) | Get A Quote |
| Gene Synthesis | Get A Quote |
Targeted protein degradation (TPD) by PROTAC (proteolysis-targeting chimera) and molecular glue small molecules is an emerging therapeutic strategy. To expand the roster of E3 ligases that can be utilized for TPD, we describe the discovery and biochemical characterization of small-molecule ligands targeting the E3 ligase KLHDC2. Furthermore, we functionalize these KLHDC2-targeting ligands into KLHDC2-based BET-family and AR PROTAC degraders and demonstrate KLHDC2-dependent target-protein degradation. Additionally, we ofer insight into the assembly of the KLHDC2 E3 ligase complex. Using biochemical binding studies, X-ray crystallography and cryo-EM, we show that the KLHDC2 E3 ligase assembles into a dynamic tet... More
