Fig. 3: The benzbromarone-binding site of hURAT1.
From: Mechanisms of urate transport and uricosuric drugs inhibition in human URAT1
a A cut-open view of the benzbromarone-binding site of hURAT1. The surface of hURAT1 is colored by the electrostatic potential calculated using Pymol. Benzbromarone is shown as yellow spheres. b Electron densities of benzbromarone and nearby residues. The map is shown as grey surface and contoured at 8.2 σ. Benzbromarone is shown as yellow sticks. The nearby residues are colored the same as in Fig. 1f. c Interactions between benzbromarone and hURAT1. Benzbromarone and its interacting residues are shown as sticks. Putative polar interactions are depicted as black dashed lines. d Urate uptake activities of various mutants of hURAT1. Data are normalized to hURAT1WT, shown as means ± s.d.; n = 3 technical replicates. The experiment was performed independently twice with similar results. Statistical significance compared with hURAT1WT was determined using ordinary one-way ANOVA with Dunnett’s multiple comparisons test. ****P < 0.0001; NS, not significant. P value between hURAT1WT and F360T, F364Y, and N237A mutants are 0.2156, 0.2114 and 0.1007, respectively. e Surface expression of various mutants of hURAT1. Data are normalized to hURAT1WT, shown as means ± s.d.; n = 3 technical replicates. The experiment was performed independently three times with similar results. Statistical significance compared with hURAT1WT was determined using ordinary one-way ANOVA with Dunnett’s multiple comparisons test. ****P < 0.0001; NS, not significant. P value of normalized surface expression between hURAT1WT and the mutant F360T is 0.1071. f Dose-dependent inhibition of hURAT1WT and various mutants by benzbromarone. Data are shown as means ± s.d.; n = 3 technical replicates. The experiment was performed independently twice with similar results. Data for hURAT1WT is the same as shown in Fig. 1d.
