4AO5

B. subtilis prophage dUTPase YosS in complex with dUMP

4AO5 の概要

• エントリーDOI: 10.2210/pdb4ao5/pdb
• 関連するPDBエントリー: 2BAZ 2XX6 2XY3 2Y1T
• 分子名称: SPBC2 PROPHAGE-DERIVED DEOXYURIDINE 5'-TRIPHOSPHATE NUCLEO TIDOHYDROLASE YOSS, 2'-DEOXYURIDINE 5'-MONOPHOSPHATE, SODIUM ION, ... (4 entities in total)
• 機能のキーワード: hydrolase, homotrimeric dutpases, phe-lid
• 由来する生物種: BACILLUS SUBTILIS
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 100350.58

構造登録者

Garcia-Nafria, J.,Harrison, C.,Turkenburg, J.P.,Wilson, K.S. (登録日: 2012-03-23, 公開日: 2013-04-03, 最終更新日: 2023-12-20)

主引用文献

Garcia-Nafria, J.,Timm, J.,Harrison, C.,Turkenburg, J.P.,Wilson, K.S.
Tying Down the Arm in Bacillus Dutpase: Structure and Mechanism
Acta Crystallogr.,Sect.D, 69:1367-, 2013

PubMed Abstract: Homotrimeric dUTPases contain three active sites, each formed by five conserved sequence motifs originating from all three subunits. The essential fifth motif lies in a flexible C-terminal arm which becomes ordered during catalysis and is disordered in most crystal structures. Previously, it has been shown that the two Bacillus subtilis dUTPases, YncF and YosS, differ from their orthologues in the position in the sequence of the essential Phe-lid residue, which stacks against the uracil base, and in the conformation of the general base aspartate, which points away from the active site. Here, three structures of the complex of YncF with dU-PPi-Mg(2+) and the structure of YosS complexed with dUMP are reported. dU-PPi-Mg(2+) triggers the ordering of both the C-terminal arm and a loop (residues 18-26) which is uniquely disordered in the Bacillus dUTPases. The dUMP complex suggests two stages in substrate release. Limited proteolysis experiments allowed those complexes in which C-terminal cleavage is hindered and those in which it can be assumed to be ordered to be identified. The results lead to the suggestion that dUpNHpp is not a perfect substrate mimic, at least for the B. subtilis enzymes, and provide new insights into the mechanism of these two dUTPases in comparison to their orthologues. The enzyme mechanism is reviewed using the present and previous crystal structures as snapshots along the reaction coordinate.

PubMed: 23897460
DOI: 10.1107/S090744491300735X

実験手法

X-RAY DIFFRACTION (1.6 Å)