Breaking antimicrobial resistance by disrupting extracytoplasmic protein folding.
Breaking antimicrobial resistance by disrupting extracytoplasmic protein folding.
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DOI：
10.7554/elife.57974
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发表时间：
2022-01-13
期刊：
影响因子：
7.7
通讯作者：
Mavridou DAI
中科院分区：
文献类型：
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作者：
Furniss RCD;Kaderabkova N;Barker D;Bernal P;Maslova E;Antwi AAA;McNeil HE;Pugh HL;Dortet L;Blair JMA;Larrouy-Maumus G;McCarthy RR;Gonzalez D;Mavridou DAI
革兰氏阴性菌的耐药性是对全球健康的最大威胁之一。迫切需要新的抗菌策略，并且开发中和抗性蛋白或损害细胞包膜完整性的抗生素佐剂越来越受到关注。大多数可用的佐剂仅对特异性抗性蛋白有效。在这里，我们表明，破坏细胞包膜蛋白的稳态同时妥协的几类阻力决定因素。特别是，我们发现，损害DsbA介导的二硫键形成使不同的β-内酰胺酶失能，并使移动的粘菌素耐药酶不稳定。此外，我们表明，DsbA的化学抑制敏感的多药耐药临床分离株现有的抗生素和DsbA的情况下，与抗生素治疗相结合，大大增加了大蜡螟幼虫感染的多药耐药铜绿假单胞菌的生存。这项工作为开发新型抗生素佐剂奠定了基础，这些佐剂可作为广泛作用的耐药性破坏剂。抗生素，如青霉素，是现代医学的基础，但细菌正在进化以抵抗它们的作用。一些最有害的病原体属于一组称为“革兰氏阴性细菌”，它们有一个外层-称为细胞包膜-作为药物屏障。这种包膜含有抗生素抗性蛋白，可以使抗生素失活或排斥抗生素，甚至在抗生素进入细胞后将其泵出细胞。解决抗生素耐药性的一种方法可能是阻止这些蛋白质发挥作用。蛋白质是由氨基酸组成的长链，可以折叠成特定的形状。为了使蛋白质正确发挥作用，它必须以正确的方式折叠。在细菌中，一种名为DsbA的蛋白质通过将其他蛋白质固定在适当的位置并插入称为二硫键的连接来帮助它们正确折叠。目前还不清楚DsbA是否在抗生素耐药性蛋白的折叠中发挥作用，但如果它确实发挥作用，它可能会开辟治疗抗生素耐药性感染的新方法。为了找到更多的答案，Kaderabkova等人收集了编码几种抗生素耐药蛋白的基因，并将它们放入大肠杆菌细菌中，使细菌对抗生素产生耐药性。Kaderabkova等人随后停止了改良的E.这导致抗生素抗性蛋白变得不稳定并分解，因为它们不能正确折叠。进一步的实验表明，在革兰氏阴性菌的其他致病物种中用化学抑制剂阻断DsbA，使这些细菌对它们通常会抵抗的抗生素更敏感。为了证明使用这种方法可以阻止这些细菌的感染，Kaderabkova等人使用了产生抗生素抗性蛋白但不能使DsbA感染昆虫幼虫的革兰氏阴性细菌。然后用抗生素处理幼虫，这提高了它们的存活率，这表明阻断DsbA可能是解决抗生素耐药细菌的好方法。根据世界卫生组织的说法，开发针对革兰氏阴性菌的新疗法至关重要，但新药的发现已经停止。解决这个问题的一个方法是开发使现有药物更好地发挥作用的方法。制造阻断DsbA的药物可以提供一种在未来使用现有抗生素治疗耐药感染的方法。
Antimicrobial resistance in Gram-negative bacteria is one of the greatest threats to global health. New antibacterial strategies are urgently needed, and the development of antibiotic adjuvants that either neutralize resistance proteins or compromise the integrity of the cell envelope is of ever-growing interest. Most available adjuvants are only effective against specific resistance proteins. Here, we demonstrate that disruption of cell envelope protein homeostasis simultaneously compromises several classes of resistance determinants. In particular, we find that impairing DsbA-mediated disulfide bond formation incapacitates diverse β-lactamases and destabilizes mobile colistin resistance enzymes. Furthermore, we show that chemical inhibition of DsbA sensitizes multidrug-resistant clinical isolates to existing antibiotics and that the absence of DsbA, in combination with antibiotic treatment, substantially increases the survival of Galleria mellonella larvae infected with multidrug-resistant Pseudomonas aeruginosa. This work lays the foundation for the development of novel antibiotic adjuvants that function as broad-acting resistance breakers. Antibiotics, like penicillin, are the foundation of modern medicine, but bacteria are evolving to resist their effects. Some of the most harmful pathogens belong to a group called the 'Gram-negative bacteria', which have an outer layer – called the cell envelope – that acts as a drug barrier. This envelope contains antibiotic resistance proteins that can deactivate or repel antibiotics or even pump them out of the cell once they get in. One way to tackle antibiotic resistance could be to stop these proteins from working. Proteins are long chains of building blocks called amino acids that fold into specific shapes. In order for a protein to perform its role correctly, it must fold in the right way. In bacteria, a protein called DsbA helps other proteins fold correctly by holding them in place and inserting links called disulfide bonds. It was unclear whether DsbA plays a role in the folding of antibiotic resistance proteins, but if it did, it might open up new ways to treat antibiotic resistant infections. To find out more, Furniss, Kaderabkova et al. collected the genes that code for several antibiotic resistance proteins and put them into Escherichia coli bacteria, which made the bacteria resistant to antibiotics. Furniss, Kaderabkova et al. then stopped the modified E. coli from making DsbA, which led to the antibiotic resistance proteins becoming unstable and breaking down because they could not fold correctly. Further experiments showed that blocking DsbA with a chemical inhibitor in other pathogenic species of Gram-negative bacteria made these bacteria more sensitive to antibiotics that they would normally resist. To demonstrate that using this approach could work to stop infections by these bacteria, Furniss, Kaderabkova et al. used Gram-negative bacteria that produced antibiotic resistance proteins but could not make DsbA to infect insect larvae. The larvae were then treated with antibiotics, which increased their survival rate, indicating that blocking DsbA may be a good approach to tackling antibiotic resistant bacteria. According to the World Health Organization, developing new treatments against Gram-negative bacteria is of critical importance, but the discovery of new drugs has ground to a halt. One way around this is to develop ways to make existing drugs work better. Making drugs that block DsbA could offer a way to treat resistant infections using existing antibiotics in the future.