Exploiting Differences in Heme Biosynthesis between Bacterial Species to Screen for Novel Antimicrobials.
Exploiting Differences in Heme Biosynthesis between Bacterial Species to Screen for Novel Antimicrobials.
复制标题
利用细菌物种之间血红素生物合成的差异，以筛选新的抗菌剂。
DOI：
10.3390/biom13101485
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发表时间：
2023-10-06
期刊：
影响因子：
5.5
通讯作者：
中科院分区：
文献类型：
--
作者：
血红素生物合成的最后三个步骤在双层和单层细菌之间表现出显着的差异。前者采用原卟啉依赖性（PPD）途径，而后者利用最近发现的粪卟啉依赖性（CPD）途径。为了快速筛选区分这两种途径的潜在抑制剂，采用滑动模块化基因置换重组工程策略，将大肠杆菌YFP菌株中与原卟啉途径相关的基因替换为金黄色葡萄球菌（Staphylococcus aureus，SA）中CPD途径相关的基因，以产生大肠杆菌YFP菌株。coli Sa-CPD-YFP。通过筛选针对产生YFP的Sa-CPD-YFP菌株与产生CFP的E. coli菌株。使用混合菌株测定，通过一种荧光信号的减少而不是另一种荧光信号的减少来鉴定靶向CPD或PPD血红素途径的抑制剂。初步筛选鉴定了唑类和灵菌红素衍生的化合物，这些化合物显示特异性靶向CPD途径并导致粪血红素的积累，这表明抑制的主要靶标似乎是粪血红素脱羧酶（ChdC）。计算机模拟强调，这些抑制剂能够结合在ChdC的活性位点内。
The final three steps of heme biogenesis exhibit notable differences between di- and mono-derm bacteria. The former employs the protoporphyrin-dependent (PPD) pathway, while the latter utilizes the more recently uncovered coproporphyrin-dependent (CPD) pathway. In order to devise a rapid screen for potential inhibitors that differentiate the two pathways, the genes associated with the protoporphyrin pathway in an Escherichia coli YFP strain were replaced with those for the CPD pathway from Staphylococcus aureus (SA) through a sliding modular gene replacement recombineering strategy to generate the E. coli strain Sa-CPD-YFP. Potential inhibitors that differentially target the pathways were identified by screening compound libraries against the YFP-producing Sa-CPD-YFP strain in comparison to a CFP-producing E. coli strain. Using a mixed strain assay, inhibitors targeting either the CPD or PPD heme pathways were identified through a decrease in one fluorescent signal but not the other. An initial screen identified both azole and prodigiosin-derived compounds that were shown to specifically target the CPD pathway and which led to the accumulation of coproheme, indicating that the main target of inhibition would appear to be the coproheme decarboxylase (ChdC) enzyme. In silico modeling highlighted that these inhibitors are able to bind within the active site of ChdC.
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影响因子：
3.7
作者：
Chait R;Shrestha S;Shah AK;Michel JB;Kishony R
通讯作者：
Kishony R
影响因子：
3.7
作者：
Clifford RJ;Milillo M;Prestwood J;Quintero R;Zurawski DV;Kwak YI;Waterman PE;Lesho EP;Mc Gann P
通讯作者：
Mc Gann P
影响因子：
6.4
作者：
Hammer ND;Reniere ML;Cassat JE;Zhang Y;Hirsch AO;Indriati Hood M;Skaar EP
通讯作者：
Skaar EP
影响因子：
3.2
作者：
Parish, T;Schaeffer, M;Duncan, K
通讯作者：
Duncan, K
影响因子：
14.9
作者：
Li XT;Thomason LC;Sawitzke JA;Costantino N;Court DL
通讯作者：
Court DL