5hx1 | pdb_00005hx1

Crystal structure of DR2231_E46A mutant in complex with dUMP and magnesium

spinqualitylabelsall models 5hx1, resolution 1.80Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5hx1 is a 4 chain structure with sequence from Deinococcus radiodurans. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.799Å Ligands: MG, UMP Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function Q9RS96_DEIRA Publication Abstract from PubMed DR2231 from Deinococcus radiodurans was previously functionally and structurally characterized as an all-alpha NTP pyrophosphohydrolase with specific dUTPase activity. dUTPases have a central role in the regulation of dUTP intracellular levels and dTTP nucleotide metabolism. DR2231 presents a conserved di-metal catalytic site, similar to the all-alpha dimeric dUTPases, but contrary to these enzymes, it is unable to process dUDP. In this article we present functional and structural evidence of single-point mutations that affect directly or indirectly the enzyme catalysis and provide a complete description of the all-alpha NTP pyrophosphohydrolase mechanism. Activity assays, isothermal titration calorimetry and the crystal structures of these mutants obtained in complex with dUMP or a dUTP analogue aid in probing the reaction mechanism. Our results demonstrate that the two metals are necessary for enzyme processing and also important to modulate the substrate binding affinity. Single-point mutations located in a structurally mobile lid-like loop show that the interactions with the nucleoside monophosphate are essential for induction of the closed conformation and ultimately for substrate processing. beta and gamma-phosphates are held in place through coordination with the second metal, which is responsible for the substrate "gauche" orientation in the catalytic position. The lack of sufficient contacts to orient the dUDP beta-phosphate for hydrolysis explains DR2231 preference towards dUTP. Sequence and structural similarities with MazG proteins suggest that a similar mechanism might be conserved within the protein family. This article is protected by copyright. All rights reserved. Deinococcus radiodurans DR2231 is a two-metal-ion mechanism hydrolase with exclusive activity on dUTP.,Mota CS, Goncalves AM, de Sanctis D FEBS J. 2016 Oct 14. doi: 10.1111/febs.13923. PMID:27739259 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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