Figure 2 |. Comparison of prospective screens against the crystal and AF2 structures of the σ₂ receptor.

a. The same 490 million molecules from ZINC20 were screened against both the crystal and AF2 structures of the σ₂ receptor. From these, 138 molecules from the crystal docking campaign and 119 from the AF2 docking campaign were synthesized and tested in a radioligand displacement assay. The campaign involving the crystal structure has already been published. The left panel is replotted based on the previously published data set. Displacement of the radioligand [³H]-DTG by each tested molecule occurs at 1 μM (mean ± s.e.m. of three technical replicates). A dashed line indicates 50% radioligand displacement. Dots below the dashed line represent confirmed binders, which are colored in blue. b. The distribution of binding affinity levels among the hits from both the AF2 and crystal structure screens. We measured competition binding curves for the 21 top docking hits from the crystal structure screen and the 18 top hits from the AF2 structure screen. These hits are categorized into three affinity ranges: <5 nM; 5 nM–50 nM; and >50 nM. c. The docked poses of the best binder from the screen against the AF2 model. d. The competition binding curve of the best binder from c against the σ₂ receptor. The data are represented as mean ± s.e.m. from three technical replicates.