9win

Crystal structure of the human dihydroorotase domain complexed with 5-fluoroorotic acid

spinqualitylabelsall models 9win, resolution 1.55Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 9win is a 1 chain structure with sequence from Homo sapiens. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.55Å Ligands: FOT, KCX, ZN Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function PYR1_HUMAN This protein is a "fusion" protein encoding four enzymatic activities of the pyrimidine pathway (GATase, CPSase, ATCase and DHOase). Publication Abstract from PubMed Dihydroorotase (DHOase) catalyzes the reversible conversion of N-carbamoyl-L-aspartate to dihydroorotate in de novo pyrimidine biosynthesis. In humans, DHOase (huDHOase) is part of the CAD enzyme complex and contains a flexible loop that alternates between loop-in and loop-out conformations. Here, we identify 5-aminoorotic acid (5-AOA), previously known as a dihydroorotate dehydrogenase inhibitor, as a potent huDHOase inhibitor. Enzyme assays showed strong inhibition by 5-AOA (IC(50) = 9.87 muM) compared with weaker inhibition by 5-fluoroorotic acid (5-FOA; IC(50) = 191.59 muM). To elucidate the mechanism, we determined the crystal structures of huDHOase bound to 5-AOA (1.83 A; PDB ID: 9WIC) and to 5-FOA (1.55 A; PDB ID: 9WIN) for direct comparison. Structural analysis revealed distinct binding modes: 5-AOA bound in the loop-in conformation, forming extensive stabilizing interactions, whereas 5-FOA bound in the loop-out state with fewer contacts. Consistently, the T1562A mutation decreased the binding affinity of 5-AOA from 18.6 muM to 53.4 muM, an approximately threefold reduction, confirming the role of the loop in inhibitor recognition. These findings establish 5-AOA as a dual inhibitor in pyrimidine biosynthesis and highlight a loop-in binding mechanism for huDHOase inhibition. Structural basis of potent inhibition of the human CAD dihydroorotase domain by 5-aminoorotic acid.,Huang YH, Huang TY, Wang MC, Huang CY Biochem Biophys Res Commun. 2025 Nov 1;787:152804. doi: , 10.1016/j.bbrc.2025.152804. Epub 2025 Oct 13. PMID:41101239 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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