2WE1

EBV dUTPase mutant Asp131Asn with bound dUMP

2WE1 の概要

• エントリーDOI: 10.2210/pdb2we1/pdb
• 関連するPDBエントリー: 2BSY 2BT1 2WE0 2WE2 2WE3
• 分子名称: DEOXYURIDINE 5'-TRIPHOSPHATE NUCLEOTIDOHYDROLASE, 2'-DEOXYURIDINE 5'-MONOPHOSPHATE, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: pyrophosphatase, epstein-barr virus, nucleotide metabolism, dutpase, monomer, hydrolase, herpes virus
• 由来する生物種: HUMAN HERPESVIRUS 4 (EPSTEIN-BARR VIRUS)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 32260.70

構造登録者

Freeman, L.,Buisson, M.,Tarbouriech, N.,Burmeister, W.P. (登録日: 2009-03-27, 公開日: 2009-07-07, 最終更新日: 2024-10-16)

主引用文献

Freeman, L.,Buisson, M.,Tarbouriech, N.,Van Der Heyden, A.,Labbe, P.,Burmeister, W.P.
The Flexible Motif V of Epstein-Barr Virus Deoxyuridine 5'-Triphosphate Pyrophosphatase is Essential for Catalysis.
J.Biol.Chem., 284:25280-, 2009

PubMed Abstract: Deoxyuridine 5'-triphosphate pyrophosphatases (dUTPases) are ubiquitous enzymes essential for hydrolysis of dUTP, thus preventing its incorporation into DNA. Although Epstein-Barr virus (EBV) dUTPase is monomeric, it has a high degree of similarity with the more frequent trimeric form of the enzyme. In both cases, the active site is composed of five conserved sequence motifs. Structural and functional studies of mutants based on the structure of EBV dUTPase gave new insight into the mechanism of the enzyme. A first mutant allowed us to exclude a role in enzymatic activity for the disulfide bridge involving the beginning of the disordered C terminus. Sequence alignments revealed two groups of dUTPases, based on the position in sequence of a conserved aspartic acid residue close to the active site. Single mutants of this residue in EBV dUTPase showed a highly impaired catalytic activity, which could be partially restored by a second mutation, making EBV dUTPase more similar to the second group of enzymes. Deletion of the flexible C-terminal tail carrying motif V resulted in a protein completely devoid of enzymatic activity, crystallizing with unhydrolyzed Mg(2+)-dUTP complex in the active site. Point mutations inside motif V highlighted the essential role of lid residue Phe(273). Magnesium appears to play a role mainly in substrate binding, since in absence of Mg(2+), the K(m) of the enzyme is reduced, whereas the k(cat) is less affected.

PubMed: 19586911
DOI: 10.1074/JBC.M109.019315

実験手法

X-RAY DIFFRACTION (1.8 Å)