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. 2022 Jun 23;13:3586. doi: 10.1038/s41467-022-31206-6

Fig. 3. Macrocycle stabilization of the 14-3-3/CFTR interaction.

Fig. 3

a Chemical structure of CY0072424. b Electron density map (unbiased Fo-Fc, 2.5σ, directly after molecular replacement) of CY007424 bound to the interface of 14-3-3β and CFTRpS753. c Final 2Fo-Fc electron density map (1σ). d Crystal structure of the 14-3-3β homodimer complexed with CFTRpS753pS768 and CY007424 (PDB ID: 7QI1). e Overlay of the CFTRpS753 peptide motif bound to 14-3-3 in the presence of FC-A (cyan sticks) or CY007424 (green sticks). FC-A is shown as purple sticks. f FP assay of FITC-labeled CFTRpS753pS768 (10 nM) with 14-3-3β in the presence of 100 μM of FC-A, CY007424, or both compounds. Background polarization was subtracted from all values, n = 3 technical replicates. g Immunoprecipitation study to show the effect of CY007424 on the interaction between of CFTR and 14-3-3 proteins. CFTR and separately 14-3-3 immunoprecipitation from HEK293 cells expressing F508del-CFTR after 24 h of CY007424 (10μM) (424) or vehicle alone (CON). LYS: cellular lysate prior to preclear, SUP: supernatant after specific bead incubation, PC: second preclear, LW: last wash after the immunoprecipitation IP are the specific antibody beads. IP, upper blot: probed with CFTR AB, lower blot: probed with 14-3-3 AB. Source data are provided as a Source Data file.