Identification of Novel Inhibitors of Escherichia coli DNA Ligase (LigA).
Identification of Novel Inhibitors of Escherichia coli DNA Ligase (LigA).
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DOI：
10.3390/molecules26092508
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发表时间：
2021-04-25
期刊：
影响因子：
--
通讯作者：
Gowers DM
中科院分区：
文献类型：
--
作者：
Alomari A;Gowland R;Southwood C;Barrow J;Bentley Z;Calvin-Nelson J;Kaminski A;LeFevre M;Callaghan AJ;Vincent HA;Gowers DM
DNA连接酶存在于所有生物体中，催化3′羟基和5′磷酸之间形成磷酸二酯键，这是在复制和修复过程中维持基因组完整性所必需的反应。真细菌DNA连接酶使用NAD+作为辅因子，并且相对于使用ATP作为辅因子的真核DNA连接酶具有低序列和结构同源性。这些关键差异使得能够特异性靶向细菌DNA连接酶作为抗菌策略。在这项研究中，四个小分子的功能重要的区域内的大肠杆菌连接酶（EC-LigA）的可访问的网站进行了鉴定，使用计算机模拟的方法。然后使用分子对接来筛选预测与这些位点结合的小分子。然后在体外连接酶测定中筛选8种候选抑制剂的抑制活性。其中5种（遗传霉素、氯己定、谷胱甘肽（还原）、咪唑烷基脲和2-（氨甲基）咪唑）显示出对EC-LigA的剂量依赖性抑制，半数最大抑制浓度（IC 50）在微摩尔至毫摩尔范围内（11-2600 µM）。两个（遗传霉素和洗必泰）被预测为结合到一个区域的EC-LigA，以前没有直接研究，提高了可能性，可能有氨基酸在这个区域内是重要的EC-LigA活性或功能的必需残基接近这个区域的影响抑制剂与这个区域的相互作用。我们预计，所确定的小分子结合位点和抑制剂可以作为靶向细菌DNA连接酶的抗菌策略的一部分。
Present in all organisms, DNA ligases catalyse the formation of a phosphodiester bond between a 3′ hydroxyl and a 5′ phosphate, a reaction that is essential for maintaining genome integrity during replication and repair. Eubacterial DNA ligases use NAD+ as a cofactor and possess low sequence and structural homology relative to eukaryotic DNA ligases which use ATP as a cofactor. These key differences enable specific targeting of bacterial DNA ligases as an antibacterial strategy. In this study, four small molecule accessible sites within functionally important regions of Escherichia coli ligase (EC-LigA) were identified using in silico methods. Molecular docking was then used to screen for small molecules predicted to bind to these sites. Eight candidate inhibitors were then screened for inhibitory activity in an in vitro ligase assay. Five of these (geneticin, chlorhexidine, glutathione (reduced), imidazolidinyl urea and 2-(aminomethyl)imidazole) showed dose-dependent inhibition of EC-LigA with half maximal inhibitory concentrations (IC50) in the micromolar to millimolar range (11–2600 µM). Two (geneticin and chlorhexidine) were predicted to bind to a region of EC-LigA that has not been directly investigated previously, raising the possibility that there may be amino acids within this region that are important for EC-LigA activity or that the function of essential residues proximal to this region are impacted by inhibitor interactions with this region. We anticipate that the identified small molecule binding sites and inhibitors could be pursued as part of an antibacterial strategy targeting bacterial DNA ligases.
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DOI：
10.1073/pnas.57.5.1426
发表时间：
1967-01-01
影响因子：
11.1
作者：
OLIVERA, BM;LEHMAN, IR
通讯作者：
LEHMAN, IR
影响因子：
4.9
作者：
Ciarrocchi, G;MacPhee, DG;Tilley, L
通讯作者：
Tilley, L
影响因子：
2.9
作者：
GRIFFITH, OW
通讯作者：
GRIFFITH, OW
DOI：
10.1007/bf00330179
发表时间：
1986-01-01
期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
ISHINO, Y;SHINAGAWA, H;NAKATA, A
通讯作者：
NAKATA, A
影响因子：
3.9
作者：
Aslam B;Wang W;Arshad MI;Khurshid M;Muzammil S;Rasool MH;Nisar MA;Alvi RF;Aslam MA;Qamar MU;Salamat MKF;Baloch Z
通讯作者：
Baloch Z