Fig. 4: Activity of Homo-BacPROTACs in cell-free degradation assays.
From: Homo-BacPROTAC-induced degradation of ClpC1 as a strategy against drug-resistant mycobacteria
a Degradation curves of ClpC1-NTD in the cell-free degradation assay (quantified by capillary Western (WES)) induced by Homo-BacPROTAC 8 (UdSBI-0545) compared to its enantiomer 8a (UdSBI-0966), matching monomer 5 and dCymC (3 independent experiments done in triplicates). b WES visualization of ClpC1-NTD degradation after titration of Homo-BacPROTACs, developed with anti-His antibody recognizing His6-tagged ClpC1-NTD and processed, His4-tagged ClpP1P2. Concentration-dependent degradation of ClpC1-NTD can be observed for 8 (UdSBI-0545) (lanes 10–13) but not for 8a (UdSBI-0966) (lanes 2–8). c Analogous to a, except that exit vector 7-based Homo-BacPROTAC 12 (UdSBI-4377), enantiomer 12a (UdSBI-0117) and monomer 10 were used. d WES-derived gel picture visualizing ClpC1-NTD degradation (lane 2–5) from representative experiment summarized in c. e SYPROTM Ruby-stained SDS-PAGE gel from exemplary cell-free degradation assay depicting efficient degradation (lane 3,4) of ClpC1-NTD by Homo-BacPROTAC 8 (UdSBI-0545), while full length ClpC1 is not significantly degraded. Green vertical lines indicate the DC50 (for 8, 12). Error bars represent mean ± SD of n = 3 independent experiments in triplicates. The actual mean DC50 values for all cell-free degradation experiments conducted for this study are summarized in Supplementary Table 8. Source data are provided as a Source Data file.
