An Azobenzene G-quadruplex Ligand Exhibits Promising Antibacterial Activity against Escherichia coli
An Azobenzene G-quadruplex Ligand Exhibits Promising Antibacterial Activity against Escherichia coli
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偶氮苯 G-四联体配体对大肠杆菌表现出良好的抗菌活性
DOI：
10.1101/2022.09.01.506212
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Ramos-Soriano J
中科院分区：
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作者：
Ramos-Soriano J
随着细菌抗菌素耐药性（AMR）的上升和抗生素发现量的下降，全球医疗保健正处于危险之中。我们非常需要开发新型抗菌剂，针对对现有抗生素产生耐药性的细菌，特别是革兰氏阴性菌，如大肠杆菌，它会导致已经受损的患者出现机会性感染。在这里，我们证明了一种新的吡啶功能化偶氮苯支架dl20，被确定为细菌基因组/转录组中的候选配体靶g -四重体（G4）结构，对多种耐药药物具有良好的抗菌活性（MIC值≤4 μ g/ml）。杆菌。串联质量标签（TMT）蛋白质组学在大肠杆菌培养中的应用。用亚致死浓度的fl20处理大肠型菌株ATCC 25922，鉴定出7个含g4的序列作为l20的潜在靶点。FRET（荧光共振能量转移）稳定实验表明，与G4的两个比较配体（刚性-二苯乙烯el5和pyridostatin(PDS)）相比，atel20以可变和中等的亲和力结合这些选定的序列，这两个比较配体（刚性-二苯乙烯el5和pyridostatin(PDS)）更好地稳定了G4结构，但表现出较低的抗菌活性。然而，蛋白质组学实验也表明，除了其优越的抗菌活性外，l20处理比l5或PDS影响更多g4相关蛋白的表达水平，并上调参与翻译的多种必需蛋白。这些发现确定了发现潜在G4配体的策略，这些方法可以导致抗菌候选物对多重耐药等优先靶点具有活性。针对G4序列和配体如l20，值得进一步探索作为G4介导作用模式的潜在新疗法。
With the rise in bacterial antimicrobial resistance (AMR) and decline in antibiotic discovery, global healthcare is at a point of jeopardy. There is great need for the development of novel antimicrobials that target bacteria that have become resistant to our existing antibiotics, in particular Gram-negative species such asEscherichia coliwhich is responsible for opportunistic infections of already compromised patients. Here we demonstrate that a novel pyridinium-functionalised azobenzene scaffoldL20, identified as a candidate ligand able target G-quadruplex (G4) structures in bacterial genomes/transcriptomes, shows promising antibacterial activity (MIC values ≤ 4 µg/ml) against multi-drug resistantE. coli. Tandem Mass Tag (TMT) proteomics applied to cultures of theE. colitype strain ATCC 25922 treated with sub-lethal concentrations ofL20, identified seven G4-containing sequences as potential targets forL20. FRET (fluorescence resonance energy transfer) stabilisation assays indicateL20binds these selected sequences with variable and moderate affinity, in contrast to two comparator G4 ligands (stiff-stilbeneL5andpyridostatin(PDS)) that better stabilise G4 structures but exhibit a lower antimicrobial activity. However, proteomic experiments also reveal that, alongside its superior antibacterial activity,L20treatment influences expression levels of more G4-associated proteins than eitherL5or PDS, and upregulates multiple essential proteins involved in translation. These findings identify strategies discovering potential G4 ligands as approaches that can lead to antibacterial candidates active against priority targets such as multi-drug resistantE. coli, and that targeting G4 sequences, and ligands such asL20, warrant further exploration as potential novel therapeutics with G4-mediated modes of action.