Selective inhibition of DNA replicase assembly by a non-natural nucleotide: exploiting the structural diversity of ATP-binding sites.
Selective inhibition of DNA replicase assembly by a non-natural nucleotide: exploiting the structural diversity of ATP-binding sites.
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DOI：
10.1021/cb900218c
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发表时间：
2010-02-19
影响因子：
4
通讯作者：
Berdis, Anthony
中科院分区：
文献类型：
--
作者：
Eng, Kevin;Scouten-Ponticelli, Sarah K.;Sutton, Mark;Berdis, Anthony
DNA合成是由一系列被称为复制酶的蛋白质催化的。这种多蛋白质复合物的有效组装对于DNA复制的连续性是必不可少的，并且由使用ATP结合和水解来协调这些事件的夹加载辅助蛋白介导。因此，选择性抑制这些辅助蛋白的活性的能力提供了一种合理的方法来调节DNA合成。为了实现这一目标，我们测试了几种非天然核苷酸抑制与DNA复制酶组装相关的ATP依赖性酶的能力。动力学和生物物理学研究鉴定了5-硝基-吲哚基-2 '-脱氧核糖-5'-三磷酸作为一种独特的非天然核苷酸，其能够选择性抑制噬菌体T4夹装载剂而不是来自大肠杆菌的同源酶。建模研究突出了每种酶的ATP结合位点之间的结构多样性，并提供了一种解释各种取代吲哚基-2 '-脱氧核糖-5'-三磷酸效力差异的机制。测量噬斑形成的体内测定证明了5-硝基-吲哚基-2 '-脱氧核糖作为细胞生长抑制剂对T4噬菌体的效力和选择性，同时保留E.大肠杆菌宿主不受影响。该策略提供了一种新的方法来开发选择性抑制有效DNA复制所需的ATP依赖性酶的试剂。
DNA synthesis is catalyzed by an ensemble of proteins designated the replicase. The efficient assembly of this multi-protein complex is essential for the continuity of DNA replication and is mediated by clamp-loading accessory proteins that use ATP binding and hydrolysis to coordinate these events. As a consequence, the ability to selectively inhibit the activity of these accessory proteins provides a rational approach to regulate DNA synthesis. Toward this goal, we tested the ability of several non-natural nucleotides to inhibit ATP-dependent enzymes associated with DNA replicase assembly. Kinetic and biophysical studies identified 5-nitro-indolyl-2'-deoxyribose-5'-triphosphate as a unique non-natural nucleotide capable of selectively inhibiting the bacteriophage T4 clamp loader versus the homologous enzyme from Escherichia coli. Modeling studies highlight the structural diversity between the ATP-binding site of each enzyme and provide a mechanism accounting for the differences in potencies for various substituted indolyl-2'-deoxyribose-5'-triphosphates. An in vivo assay measuring plaque formation demonstrates the efficacy and selectivity of 5-nitro-indolyl-2'-deoxyribose as a cytostatic agent against T4 bacteriophage while leaving viability of the E. coli host unaffected. This strategy provides a novel approach to develop agents that selectively inhibit ATP-dependent enzymes that are required for efficient DNA replication.
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影响因子：
2.9
作者：
Alley, SC;Abel-Santos, E;Benkovic, SJ
通讯作者：
Benkovic, SJ
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
2.9
作者：
Reineks, EZ;Berdis, AJ
通讯作者：
Berdis, AJ
DOI：
10.1016/0005-2787(67)90652-1
发表时间：
1967-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
HIRAGA, S;IGARASHI, K;YURA, T
通讯作者：
YURA, T
影响因子：
9.2
作者：
KABOORD, BF;BENKOVIC, SJ
通讯作者：
BENKOVIC, SJ