Virtual screening and activity evaluation of human uric acid transporter 1 (hURAT1) inhibitors

Yang, Yacong; Hu, Yu; Yao, Fengli; Yang, Jinbo; Ge, Leilei; Wang, Peng; Xu, Ximing

doi:10.1039/d2ra07193b

Cited by 4 publications

(1 citation statement)

References 38 publications

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“…The three-dimensional structure of RpUGT2 was predicted utilizing the Alphafold2 software, as previously elucidated (Yang et al 2023). Molecular docking of the avonoids with RpUGT2 structures was executed employing Schrödinger's Induced Fit Docking (IFD) module (Zhang et al 2023).…”

Section: Molecular Docking and Site-directed Mutagenesismentioning

confidence: 99%

New Glycosyltransferase RpUGT2 from Rheum palmatum involved in the flavonoid glycoside Biosynthesis

Zou,

Zhang

et al. 2024

Preprint

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Uridine diphosphate-dependent glycosyltransferases (UGTs) serve as key mediators in the glycosylation of plant metabolites, thereby inducing alterations in their physicochemical properties and bioactivities. Particularly, flavonoid glycosides are synthesized from aglycones through reactions catalyzed by typical UGTs. In our present investigation, we successfully cloned a novel UGT, RpUGT2, originating from Rheum palmatum, a highly regarded traditional Chinese herb known for its abundant genetic diversity. Sequence alignment and phylogenetic analysis indicated the close phylogenetic proximity of RpUGT2 to Vitis vinifera. The recombinant RpUGT2 protein, produced heterologously in Escherichia coli, displayed robust catalytic activity towards a spectrum of flavonoids. Mutational analyses revealed that specific amino acid sequence modifications could result in the loss of enzymatic activity in RpUGT2. This study has thus provided potential candidate genes, RpUGT2, for prospective applications in the metabolic engineering of flavonoid compounds in plants.

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Section: Molecular Docking and Site-directed Mutagenesismentioning

confidence: 99%

New Glycosyltransferase RpUGT2 from Rheum palmatum involved in the flavonoid glycoside Biosynthesis

Zou,

Zhang

et al. 2024

Preprint

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Virtual screening and biological evaluation of natural products as urate transporter 1 (URAT1) inhibitors

Du,

Yin,

Xie

et al. 2024

Journal of Biomolecular Structure and Dynamics

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Unique Binding Sites of Uricosuric Agent Dotinurad for Selective Inhibition of Renal Uric Acid Reabsorptive Transporter URAT1

Fujita,

Isozumi,

Zhu

et al. 2024

J Pharmacol Exp Ther

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Dotinurad was developed as a uricosuric agent, inhibiting urate (UA) reabsorption through the UA transporter URAT1 in the kidneys. Due to its high selectivity for URAT1 among renal UA transporters, we investigated the mechanism underlying this selectivity by identifying dotinurad binding sites specific to URAT1. Dotinurad was docked to URAT1 using AutoDock4, utilizing the AlphaFold2-predicted structure. The inhibitory effects of dotinurad on wild-type and mutated URAT1 at the predicted binding sites were assessed through URAT1-mediated [ 14 C]UA uptake in Xenopus oocytes. Nine amino acid residues in URAT1 were identified as dotinurad-binding sites.Sequence alignment with UA-transporting organic anion transporters (OATs) revealed that H142 and R487 were unique to URAT1 among renal UA-transporting OATs. For H142, IC 50 values of dotinurad increased to 62, 55, and 76 nM for mutated URAT1 (H142A, H142E, and H142R, respectively), compared to 19 nM for the wild-type, indicating that H142 contributes to URAT1-selective interaction with dotinurad. H142 was predicted to interact with the phenyl-hydroxyl group of dotinurad. The IC 50 of the hydroxyl group methylated dotinurad (F13141) was 165 μM, 8,420-fold higher than dotinurad, suggesting the interaction of H142 and the phenyl-hydroxyl group by forming a hydrogen bond. Regarding R487, URAT1-R487A exhibited a loss of activity. Interestingly, the URAT1-H142A/R487A double mutant restored UA transport activity, with the IC 50 value of dotinurad for the mutant (388 nM) significantly higher than that for H142A (73.5 nM). These results demonstrate that H142 and R487 of URAT1 determine its selectivity for dotinurad, a uniqueness observed only in URAT1 among UA-transporting OATs.

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Virtual screening and activity evaluation of human uric acid transporter 1 (hURAT1) inhibitors

Abstract: Alphafold2 was used to predict URAT1 protein structure, then the docking sites were identified, and three hit compounds were obtained through virtual screening and bioactivity verification.

Cited by 4 publications

References 38 publications

New Glycosyltransferase RpUGT2 from Rheum palmatum involved in the flavonoid glycoside Biosynthesis

New Glycosyltransferase RpUGT2 from Rheum palmatum involved in the flavonoid glycoside Biosynthesis

Virtual screening and biological evaluation of natural products as urate transporter 1 (URAT1) inhibitors

Unique Binding Sites of Uricosuric Agent Dotinurad for Selective Inhibition of Renal Uric Acid Reabsorptive Transporter URAT1

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