9LN0

A thermostable enzyme dUTPase P45

これはPDB形式変換不可エントリーです。

9LN0 の概要

• エントリーDOI: 10.2210/pdb9ln0/pdb
• 分子名称: dCTP deaminase (2 entities in total)
• 機能のキーワード: a thermostable enzyme dutpase p45, metal binding protein
• 由来する生物種: Pyrococcus furiosus DSM 3638
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 17893.75

構造登録者

Wang, Y.X.,Dong, B.J. (登録日: 2025-01-20, 公開日: 2025-11-19)

主引用文献

Dong, B.,Li, J.,Zhang, L.,Zhang, B.,Xu, B.,Ye, S.,Wang, Y.
Structural and functional characterization of thermostable dUTPase P45 from Pyrococcus furiosus with enhanced PCR efficiency.
Int.J.Biol.Macromol., 320:146110-146110, 2025

PubMed Abstract: dUTPase is a critical enzyme responsible for hydrolyzing dUTP to dUMP and pyrophosphate (PPi), thereby maintaining genomic integrity by preventing uracil misincorporation into DNA. P45, an archaeal dUTPase, enhances polymerase chain reaction (PCR) efficiency by increasing product yield and amplification duration. However, its oligomeric state and catalytic mechanism remain poorly characterized. Here, we report the crystal structures of Pyrococcus furiosus P45 in its apo form (P45-apo) and in complex with dUMP (P45-dUMP) at 2.1 Å and 2.2 Å resolution. Mutational studies identified key residues (W93, D95, T103, Y138) essential for enzymatic activity. Comparative structural analysis revealed that P45 shares a highly conserved catalytic core with trimeric dUTPases across diverse species, including archaea, bacteria, eukaryotes, viruses, and protozoa. Substrate binding induced conformational rearrangements, stabilizing β-sheet formation and active site closure. These findings elucidate the structural basis of P45's thermostability and PCR-enhancing activity, providing insights for its application in high-fidelity DNA amplification systems.

PubMed: 40680961
DOI: 10.1016/j.ijbiomac.2025.146110

実験手法

X-RAY DIFFRACTION (2.2 Å)