Fig. 2: Optimization and characterization of GPlad.

a Changes in fluorescence with the introduction of McsA in the GPlad system. Corresponding cells were induced with 20 ng/mL aTc for 6 h. b Evaluation of GPlad degradation with ClpCP expression controlled by P_j23105. Corresponding cells were treated with 20 ng/mL aTc for 6 h. c Optimization of the linker length between GP and McsB. Corresponding cells were treated with 20 ng/mL aTc for 6 h, and the mKate2 fluorescence was measured using a microplate reader. d, e. Binding interface selection (Left) and design models (Right) for GP complexes. Left: Target protein surfaces showing hydrophobic residues (red) and Patchdock-selected binding sites (yellow). Right: Designed complexes of GP^ErmB-ErmB and GP^AroK-AroK, with binding sites (yellow) and targets (gray). f WB analysis of ErmB abundance in genomic expression after 2 h of GP^ErmB-McsB induction. g Spot plating assays of E. coli harboring GPlad for ErmB under 150 μg/mL erythromycin stress for 12 h on plates. h WB analysis of AroK abundance in genomic expression after 2 h of GP^AroK-McsB induction. i Evaluation of changes in the kinase activity of AroK degradation following GPlad induction after 8 h. j Degradation assay of FGFR2 and VirB8. Corresponding cells were induced with 20 ng/mL aTc for 6 h, and protein abundance was measured by WB analysis. k Volcano plot showing fold changes in protein abundance from global proteomics analysis of E. coli cells induced with GPlad targeting mKate2 for 6 h. A two-sided unpaired t-test was performed on the relative quantitative value of each protein from the two sample groups. p < 0.01 was usually considered the threshold for significance. All experiments are presented as mean ± s.d. from three biologically independent replicates. Source data are provided as a Source Data file.