Characterization of Posttranslationally Modified Multidrug Efflux Pumps Reveals an Unexpected Link between Glycosylation and Antimicrobial Resistance.
Characterization of Posttranslationally Modified Multidrug Efflux Pumps Reveals an Unexpected Link between Glycosylation and Antimicrobial Resistance.
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翻译后修饰的多药物外排泵的特性揭示了糖基化与抗菌素耐药性之间的意想不到的联系。
DOI：
10.1128/mbio.02604-20
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发表时间：
2020-11-17
期刊：
影响因子：
6.4
通讯作者：
Wren BW
中科院分区：
文献类型：
--
作者：
Abouelhadid S;Raynes J;Bui T;Cuccui J;Wren BW
几乎所有的细菌都至少有一个糖基化系统，但这些翻译后蛋白质修饰的直接影响尚未得到解决。对几种细菌病原体的糖蛋白组范围的分析揭示了毒力因子和蛋白质组装体的一般聚糖修饰。使用空肠弯曲菌作为模式生物，我们研究了一般N-连接聚糖在革兰氏阴性菌中常见的多药外排泵中的作用。我们显示，第一次，N-连接的聚糖和多药外排泵活性之间的直接联系。在蛋白质水平上，我们证明了N-连接聚糖在增强蛋白质热稳定性和介导多药外排泵组装以促进抗菌药物耐药性方面发挥作用，突出了这种翻译后修饰在细菌生理学中的重要性。预计在具有一般蛋白糖基化系统的其他革兰氏阴性病原体中也会发现聚糖的类似作用。多重耐药细菌感染的大幅增加是当前全球的当务之急。表征细菌中抗菌素耐药性的累积努力已经证明了六个多药外排泵家族的传播，其中耐药-增殖-细胞分裂（RND）是革兰氏阴性菌中多药耐药的主要机制。RND由三部分蛋白质组装组成，并赋予对一系列无关化合物的抗性。在主要的肠道病原体空肠弯曲杆菌中，RND的三种蛋白质组分被N-连接聚糖后修饰。N-连接聚糖在C.空肠和其他细菌一直难以捉摸。在这里，我们提出了第一个详细的帐户的作用，N-连接的聚糖和N-糖基化之间的联系和耐药性的C。空肠。我们证明了N-连接聚糖在增强蛋白质热稳定性、稳定蛋白质复合物和促进蛋白质-蛋白质相互作用中的多功能作用，从而通过增强多药外排泵活性介导抗菌素耐药性。这证实了糖基化对于多药外排泵组装至关重要。我们提出了一个通用的策略，可用于研究一般的糖基化系统在弯曲杆菌属和一个潜在的目标，开发针对多重耐药病原体的抗菌药物。
Nearly all bacterial species have at least a single glycosylation system, but the direct effects of these posttranslational protein modifications are unresolved. Glycoproteome-wide analysis of several bacterial pathogens has revealed general glycan modifications of virulence factors and protein assemblies. Using Campylobacter jejuni as a model organism, we have studied the role of general N-linked glycans in the multidrug efflux pump commonly found in Gram-negative bacteria. We show, for the first time, the direct link between N-linked glycans and multidrug efflux pump activity. At the protein level, we demonstrate that N-linked glycans play a role in enhancing protein thermostability and mediating the assembly of the multidrug efflux pump to promote antimicrobial resistance, highlighting the importance of this posttranslational modification in bacterial physiology. Similar roles for glycans are expected to be found in other Gram-negative pathogens that possess general protein glycosylation systems. The substantial rise in multidrug-resistant bacterial infections is a current global imperative. Cumulative efforts to characterize antimicrobial resistance in bacteria has demonstrated the spread of six families of multidrug efflux pumps, of which resistance-nodulation-cell division (RND) is the major mechanism of multidrug resistance in Gram-negative bacteria. RND is composed of a tripartite protein assembly and confers resistance to a range of unrelated compounds. In the major enteric pathogen Campylobacter jejuni, the three protein components of RND are posttranslationally modified with N-linked glycans. The direct role of N-linked glycans in C. jejuni and other bacteria has long been elusive. Here, we present the first detailed account of the role of N-linked glycans and the link between N-glycosylation and antimicrobial resistance in C. jejuni. We demonstrate the multifunctional role of N-linked glycans in enhancing protein thermostability, stabilizing protein complexes and the promotion of protein-protein interaction, thus mediating antimicrobial resistance via enhancing multidrug efflux pump activity. This affirms that glycosylation is critical for multidrug efflux pump assembly. We present a generalized strategy that could be used to investigate general glycosylation system in Campylobacter genus and a potential target to develop antimicrobials against multidrug-resistant pathogens.