The structure of Ap4A hydrolase complexed with ATP-MgFx reveals the basis of substrate binding
The structure of Ap4A hydrolase complexed with ATP-MgFx reveals the basis of substrate binding
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DOI：
10.1016/s0969-2126(02)00696-2
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发表时间：
2002-02-01
期刊：
影响因子：
5.7
通讯作者：
Gooley, PR
中科院分区：
文献类型：
--
作者：
Fletcher, JI;Swarbrick, JD;Gooley, PR
AP（4）A水解酶是调节细胞内二核苷多磷酸浓度的营养酶，涉及一系列生物事件，包括热休克和代谢应激。我们已经证明，ATP·MgFx可以用来模拟来自狭叶羽扇豆的Ap（4）A水解酶结合位点的底物，并且与以前的底物类似物不同，它与酶的交换缓慢。与ATP·MgFx复合的酶的三维结构被解决，并显示出显着的构象变化。L.狭叶Ap（4）水解酶与先前发表的两种Nutrition酶ADP-核糖焦磷酸酶和MutT明显不同，尽管它们具有共同的折叠和活性位点残基的保守性。参与底物结合的大多数残基在来自病原菌的不对称Ap（4）A水解酶中是保守的，但在它们的人类对应物中不存在，这表明可能产生靶向细菌而非人类Ap（4）A水解酶的化合物。
Ap(4)A hydrolases are Nudix enzymes that regulate intracellular dinucleoside polyphosphate concentrations, implicating them in a range of biological events' including heat shock and metabolic stress. We have demonstrated that ATP.MgFx can be used to mimic substrates in the binding site of Ap(4)A hydrolase from Lupinus angustifolius and that, unlike previous substrate analogs, it is in slow exchange with the enzyme. The three-dimensional structure of the enzyme complexed with ATP.MgFx was solved and shows significant conformational changes. The substrate binding site of L. angustifolius Ap(4)A hydrolase differs markedly from the two previously published Nudix enzymes, ADP-ribose pyrophosphatase and MutT, despite their common fold and the conservation of active site residues. The majority of residues involved in substrate binding are conserved in asymmetrical Ap(4)A hydrolases from pathogenic bacteria, but are absent in their human counterparts, suggesting that it might be possible to generate compounds that target bacterial, but not human, Ap(4)A hydrolases.