Fig. 3: Exploration of distinct cellular activities, localization and affinities of the CDH switches.
From: A rational blueprint for the design of chemically-controlled protein switches
a Schematic representation of the CDHs utilized to control transcription regulation with the GAL4/UAS system (CDH-TFs). b CDH-TFs fold-change activity between drug treated (1 µM) versus untreated (DMSO), showing the quantification of SEAP expression after 24 h drug treatment. c Drug dose-dependent responses of CDH-TFs quantified by SEAP expression after 24 h drug treatment. d Schematic representation of the CDHs utilized to regulate surface signaling receptors (CDH-GEMS) in engineered cells. e CDH-GEMS fold-change activity between drug (1 µM) versus no drug treatment (DMSO), showing the quantification of SEAP expression after 24 h. f Drug dose-dependent responses of CDH-TFs quantified by SEAP expression after 24 h. g Summary of the LD3 mutants including computationally predicted decreases in affinity and experimental measurements using SPR. Values shown are the predicted interaction energy of LD3 mutants (ΔΔG in Rosetta energy units), the binding affinities with Bcl-XL and Bcl2 and IC50s of CDH dissociation also for Bcl-XL and Bcl2 with Drug-1 and Drug-2, respectively. h Drug dose-dependent responses in engineered cells determined for the CDH-(1-2)-GEMS with LD3 and LD3_v3. The drug receptor utilized were Bcl-XL (CDH-1) in red and Bcl2 (CDH-2) in green, CDHs with original LD3 were in circle symbol and LD3_v3 in square symbol with Bcl-XL and Bcl2. b, c, e, f, h Each data point represents the mean ± s.d. of three replicates and IC50s were computed using four-parameter nonlinear regression. c, f, h Each data point was normalized to the maximal response calculated using four-parameter nonlinear regression. b, c, e, f, h Source data are available in the source data file.
