Structural studies of the Nudix hydrolase DR1025 from Deinococcus radiodurans and its ligand complexes.

Structural studies of the Nudix hydrolase DR1025 from Deinococcus radiodurans and its ligand complexes.
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来自耐辐射奇球菌的 Nudix 水解酶 DR1025 及其配体复合物的结构研究。

DOI:
10.1016/j.jmb.2004.01.065
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发表时间:
2004
期刊:
Journal of molecular biology.
影响因子:
--
通讯作者:
Holbrook,StephenR
Holbrook,StephenR
中科院分区:
--
文献类型:
--
作者:
Ranatunga,Wasantha;Hill,EmmaE;Mooster,JanaL;Holbrook,ElizabethL;Schulze-Gahmen,Ursula;Xu,WenLian;Bessman,MauriceJ;Brenner,StevenE;Holbrook,StephenR

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我们测定了极端耐辐射菌Deinococcus radiodurans中的Nutriol水解酶DR 1025在1.4nm处的晶体结构。蛋白质通过在链之间交换N-末端片段形成缠结的同源二聚体。我们已经鉴定了Nuvis折叠的其他保守元件,包括金属结合基序、以Nuvis共有序列上游两个位置的脯氨酸为特征的扭结β链,以及参与形成底物结合口袋的N-末端延伸。还解析了在镁和GTP类似物或Ap 4A存在下结晶的DR 1025的晶体结构(均为1.6 nm分辨率)。在Ap 4A共晶体中,电子密度表明不对称水解产物ATP与酶结合。GTP类似物结合结构显示GTP与ATP几乎相同地结合。三磷酸核苷均未进一步裂解。
We have determined the crystal structure, at 1.4Å, of the Nudix hydrolase DR1025 from the extremely radiation resistant bacterium Deinococcus radiodurans. The protein forms an intertwined homodimer by exchanging N-terminal segments between chains. We have identified additional conserved elements of the Nudix fold, including the metal-binding motif, a kinked β-strand characterized by a proline two positions upstream of the Nudix consensus sequence, and participation of the N-terminal extension in the formation of the substrate-binding pocket. Crystal structures were also solved of DR1025 crystallized in the presence of magnesium and either a GTP analog or Ap4A (both at 1.6Å resolution). In the Ap4A co-crystal, the electron density indicated that the product of asymmetric hydrolysis, ATP, was bound to the enzyme. The GTP analog bound structure showed that GTP was bound almost identically as ATP. Neither nucleoside triphosphate was further cleaved.
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