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. 2019 Jun 7;8:e47172. doi: 10.7554/eLife.47172

Figure 2. Mechanism of action for select N-arylsulfindoline RORγ modulators.

(A) Representative structures of four modulators and their pharmacology. (B) Co-crystal structure solutions of SR19265 and SR19355 reveal mechanism of action. A peptide bond on SR19355 acts as a hydrogen bond donor for H479 and disrupts H479 and Y502 hydrogen bond. (C, D) Differential HDX-MS was employed to evaluate ligand dependent structural perturbations in solution. Representative deuterium build-up plots showing Helix 3 (C, H3) and Helix 12 (D, H12) dynamics. (E) fluorescence polarization results showing gradient of affinity for coactivator peptide depending on compound. (F) RORγ modulators were tested in a cell-based promotor reporter assay. Vectors encoding full length RORγ and a 5xRORE-Luciferase promotor reporter were transiently transfected into HEK293T cells. These cells were seeded into a 384 well plate and tested for dose-dependent responses to compounds.

Figure 2.

Figure 2—figure supplement 1. Electron density maps are shown for compounds of interest (related to Figures 1 and 2).

Figure 2—figure supplement 1.

2Fo-Fc densities are contoured at 1.5 σ. SR19265 and SR19355 are shown in panels (A and B). SR2211 in chain A and chain B are shown in figures (C and D).