Fig. 2: Inhibition mechanism of URAT1^EM by benzbromarone.

a The cryo-EM density of benzbromarone and surrounding residues is shown as mesh while the structure of which are presented as sticks. b Side view URAT1 in complex with benzbromarone. Electrostatic potential map of the cytoplasmic pocket is shown as a surface. URAT1^EM is dipicted in sky-blue cartoon, while benzbromarone is shown as spheres with carbon, oxygen, and bromine atoms colored in deep teal, red, and green, respectively. c Coronal section view of electrostatic surface of URAT1^BEN. Benzbromarone is represented as sphere. d Detailed view of the benzbromarone binding pocket. Surrounding residues are represented as sticks. Hydrogen bond is depicted as black dash line. e Comparative evaluation of the [¹⁴C] urate transportation activity of URAT1 mutants in relation to the URAT1^EM. Data are mean ± s.e.m. from three independent assays, each with one measurement (n = 3). Comparison of URAT1^EM with mutant constructs using an unpaired two-sided t-test showed significant differences (all P < 0.0001, denoted as ****). f Inhibition of ¹⁴C-urate transport by benzbromarone in URAT1^EM and its mutants. Data are presented as normalized mean ± s.e.m. from three biologically independent assays (n  =  3).