Supplementary Figure 1: In vitro electrophysiological and binding characterization of AM-1488 and AM-3607 through IonFlux HT and SPR.
From: Crystal structures of human glycine receptor α3 bound to a novel class of analgesic potentiators
(a) AM-1488 potentiates currents evoked by EC20 glycine in HEK293T cells stably expressing human GlyRα3β (EC50 = 0.26 ± 0.02 μM), as shown by the dose response plotted as percent of maximal evoked current amplitude (1 mM glycine). (b) AM-3607 potentiates currents evoked by EC20 glycine in HEK293T cells stably expressing human GlyRα3β (EC50 = 0.051 ± 0.016 μM), as shown by the dose response plotted as percent of maximal evoked current amplitude (1 mM glycine). All values are calculated EC50 ± s.e.m. and each data point is mean ± s.e.m. with N≥5 for (a) and (b). (c) Representative binding trace of AM-1488 to GlyRα3cryst measured by SPR with kinetic fit overlaid. (d) Representative binding trace of AM-3607 to GlyRα3cryst measured by SPR with kinetic fit overlaid. Values for KD are mean ± s.d. determined from N=3 experiments for (c) and (d). (e) Representative trace of AM-1488 not binding to GlyRα3cryst preincubated with AM-3607 measured by SPR. Trace is representative of N=4 total trials on two different flow cells.
