Perturbed structural dynamics underlie inhibition and altered efflux of the multidrug resistance pump AcrB.
Perturbed structural dynamics underlie inhibition and altered efflux of the multidrug resistance pump AcrB.
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扰动的结构动力学是多药耐药泵AcrB抑制和外排改变的基础。
DOI：
10.1038/s41467-020-19397-2
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发表时间：
2020-11-04
影响因子：
16.6
通讯作者：
Politis A
中科院分区：
文献类型：
--
作者：
Reading E;Ahdash Z;Fais C;Ricci V;Wang-Kan X;Grimsey E;Stone J;Malloci G;Lau AM;Findlay H;Konijnenberg A;Booth PJ;Ruggerone P;Vargiu AV;Piddock LJV;Politis A
耐药-增殖-分裂外排泵在细菌固有和进化的多药耐药中起关键作用。AcrB是该蛋白质家族的典型成员，从细菌中挤出多种抗菌剂。虽然高分辨率的结构存在的AcrB，其构象波动和他们的功能推定的作用在很大程度上是未知的。在这里，我们确定这些结构动力学的存在下，使用氢/氘交换质谱，分子动力学模拟和细菌易感性研究的补充基板。我们发现，外排泵抑制剂通过抑制药物结合口袋动力学而不是阻止抗生素结合来增强抗生素活性。我们还揭示了在多药耐药临床分离株中发现的药物结合口袋取代改变了转运途径的可塑性，这可以解释其改变的底物外排。我们的研究结果提供了深入了解药物输出的分子机制和抑制一个主要的多药外排泵和其动力学的指导作用。AcrB是一种典型的耐药-增殖-分裂（RND）细菌转运蛋白，赋予对多种抗生素的耐药性。HDX-MS和其他互补方法提供了对AcrB结构动力学的深入了解，并提出了药物输出和抑制这种多药耐药泵的分子机制。
Resistance–nodulation–division efflux pumps play a key role in inherent and evolved multidrug resistance in bacteria. AcrB, a prototypical member of this protein family, extrudes a wide range of antimicrobial agents out of bacteria. Although high-resolution structures exist for AcrB, its conformational fluctuations and their putative role in function are largely unknown. Here, we determine these structural dynamics in the presence of substrates using hydrogen/deuterium exchange mass spectrometry, complemented by molecular dynamics simulations, and bacterial susceptibility studies. We show that an efflux pump inhibitor potentiates antibiotic activity by restraining drug-binding pocket dynamics, rather than preventing antibiotic binding. We also reveal that a drug-binding pocket substitution discovered within a multidrug resistant clinical isolate modifies the plasticity of the transport pathway, which could explain its altered substrate efflux. Our results provide insight into the molecular mechanism of drug export and inhibition of a major multidrug efflux pump and the directive role of its dynamics. AcrB is a prototypical resistance–nodulation–division (RND) bacterial transporter, conferring resistance to a variety of antibiotics. HDX-MS and other, complementary approaches offer insight into AcrB structural dynamics and suggest the molecular mechanisms underlying drug export and inhibition of this multidrug-resistance conferring pump.
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DOI：
10.1126/science.1198542
发表时间：
2011-04-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bhabha G;Lee J;Ekiert DC;Gam J;Wilson IA;Dyson HJ;Benkovic SJ;Wright PE
通讯作者：
Wright PE
完整膜蛋白复合物的质谱法。
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
5.8
作者：
Lau, Andy M. C.;Ahdash, Zainab;Politis, Argyris
通讯作者：
Politis, Argyris
影响因子：
3.4
作者：
Chalmers MJ;Busby SA;Pascal BD;West GM;Griffin PR
通讯作者：
Griffin PR
影响因子：
4.5
作者：
Johnson, Rachel M.;Fais, Chiara;Bavro, Vassiliy N.
通讯作者：
Bavro, Vassiliy N.