4LRL

Structure of an Enterococcus Faecalis HD-domain protein complexed with dGTP and dTTP

4LRL の概要

• エントリーDOI: 10.2210/pdb4lrl/pdb
• 関連するPDBエントリー: 2O6I 3IRH
• 分子名称: HD domain protein, NICKEL (II) ION, 2'-DEOXYGUANOSINE-5'-TRIPHOSPHATE, ... (7 entities in total)
• 機能のキーワード: structural genomics, psi-2, protein structure initiative, midwest center for structural genomics, mcsg, hd domain, phosphohydrolase, dntpase, allosteric regulation, metal binding protein, hydrolase
• 由来する生物種: Enterococcus faecalis
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 227342.39

構造登録者

Vorontsov, I.I.,Minasov, G.,Shuvalova, L.,Joachimiak, A.,Anderson, W.F.,Midwest Center for Structural Genomics (MCSG) (登録日: 2013-07-19, 公開日: 2013-11-20, 最終更新日: 2023-09-20)

主引用文献

Vorontsov, I.I.,Wu, Y.,Delucia, M.,Minasov, G.,Mehrens, J.,Shuvalova, L.,Anderson, W.F.,Ahn, J.
Mechanisms of Allosteric Activation and Inhibition of the Deoxyribonucleoside Triphosphate Triphosphohydrolase from Enterococcus faecalis.
J.Biol.Chem., 289:2815-2824, 2014

PubMed Abstract: EF1143 from Enterococcus faecalis, a life-threatening pathogen that is resistant to common antibiotics, is a homo-tetrameric deoxyribonucleoside triphosphate (dNTP) triphosphohydrolase (dNTPase), converting dNTPs into the deoxyribonucleosides and triphosphate. The dNTPase activity of EF1143 is regulated by canonical dNTPs, which simultaneously act as substrates and activity modulators. Previous crystal structures of apo-EF1143 and the protein bound to both dGTP and dATP suggested allosteric regulation of its enzymatic activity by dGTP binding at four identical allosteric sites. However, whether and how other canonical dNTPs regulate the enzyme activity was not defined. Here, we present the crystal structure of EF1143 in complex with dGTP and dTTP. The new structure reveals that the tetrameric EF1143 contains four additional secondary allosteric sites adjacent to the previously identified dGTP-binding primary regulatory sites. Structural and enzyme kinetic studies indicate that dGTP binding to the first allosteric site, with nanomolar affinity, is a prerequisite for substrate docking and hydrolysis. Then, the presence of a particular dNTP in the second site either enhances or inhibits the dNTPase activity of EF1143. Our results provide the first mechanistic insight into dNTP-mediated regulation of dNTPase activity.

PubMed: 24338016
DOI: 10.1074/jbc.M113.524207

実験手法

X-RAY DIFFRACTION (2.35 Å)