Structural Basis of Peptide-Based Antimicrobial Inhibition of a Resistance-Nodulation-Cell Division Multidrug Efflux Pump.
Structural Basis of Peptide-Based Antimicrobial Inhibition of a Resistance-Nodulation-Cell Division Multidrug Efflux Pump.
复制标题
DOI：
10.1128/spectrum.02990-22
复制
发表时间：
2022-10-26
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
革兰氏阴性菌的耐药-增殖-细胞分裂（RND）家族中的细菌外排泵对许多病原体的抗生素耐药性的发展有重要贡献。在这项研究中，我们选择了淋球菌的MtrD转运蛋白，因为它是这种严格的人类病原体所拥有的唯一RND泵，可以输出多种抗菌剂，包括抗生素，胆汁盐，洗涤剂，染料和抗菌肽。使用我们以前发表的在存在或不存在结合抗生素的情况下MtrD结构的知识作为模型，以及MtrCDE输出阳离子抗菌肽的已知能力，我们假设阳离子肽可以容纳在MtrD结合位点内。此外，我们认为缺乏抗菌作用的MtrD结合肽可以使细菌对通常由MtrCDE外排泵或不同革兰氏阴性菌所拥有的其他类似RND型泵输出的抗生素敏感。我们现在报告一种新的非抗菌性环状阳离子抗菌肽，我们称之为CASP（阳离子抗菌增敏肽）的鉴定。通过单颗粒冷冻电子显微镜，我们发现CASP结合在MtrD的周质裂隙区域内，使用重叠和不同的氨基酸接触位点与另一种环状肽（粘菌素）或来源于人LL-37的线性人阳离子抗菌肽相互作用。虽然CASP不能使淋病奈瑟菌对作为RND泵底物的抗生素（新生霉素）敏感，但它可以对多种革兰氏阴性杆状细菌敏感。我们建议CASP（或未来的衍生物）可以作为抗生素治疗的某些革兰氏阴性菌感染的佐剂，以前被RND转运蛋白挫败。重要信息RND外排泵可以输出许多进入革兰氏阴性菌的抗菌剂，它们的作用可以降低抗生素的疗效，并降低对各种宿主抗菌剂的敏感性。在这里，我们确定了一个阳离子抗菌致敏肽（CASP），结合内周质裂缝的RND转运蛋白（MtrD）产生的淋病奈瑟菌。令人惊讶的是，CASP能够使杆状革兰氏阴性细菌而不是淋球菌对作为淋球菌MtrCDE外排泵底物的抗生素敏感。CASP（或其未来的衍生物）可用作佐剂来治疗RND外排导致多药耐药性的感染。
Bacterial efflux pumps in the resistance-nodulation-cell division (RND) family of Gram-negative bacteria contribute significantly to the development of antimicrobial resistance by many pathogens. In this study, we selected the MtrD transporter protein of Neisseria gonorrhoeae as it is the sole RND pump possessed by this strictly human pathogen and can export multiple antimicrobials, including antibiotics, bile salts, detergents, dyes, and antimicrobial peptides. Using knowledge from our previously published structures of MtrD in the presence or absence of bound antibiotics as a model and the known ability of MtrCDE to export cationic antimicrobial peptides, we hypothesized that cationic peptides could be accommodated within MtrD binding sites. Furthermore, we thought that MtrD-bound peptides lacking antibacterial action could sensitize bacteria to an antibiotic normally exported by the MtrCDE efflux pump or other similar RND-type pumps possessed by different Gram-negative bacteria. We now report the identification of a novel nonantimicrobial cyclic cationic antimicrobial peptide, which we termed CASP (cationic antibiotic-sensitizing peptide). By single-particle cryo-electron microscopy, we found that CASP binds within the periplasmic cleft region of MtrD using overlapping and distinct amino acid contact sites that interact with another cyclic peptide (colistin) or a linear human cationic antimicrobial peptide derived from human LL-37. While CASP could not sensitize Neisseria gonorrhoeae to an antibiotic (novobiocin) that is a substrate for RND pumps, it could do so against multiple Gram-negative, rod-shaped bacteria. We propose that CASP (or future derivatives) could serve as an adjuvant for the antibiotic treatment of certain Gram-negative infections previously thwarted by RND transporters. IMPORTANCE RND efflux pumps can export numerous antimicrobials that enter Gram-negative bacteria, and their action can reduce the efficacy of antibiotics and provide decreased susceptibility to various host antimicrobials. Here, we identified a cationic antibiotic-sensitizing peptide (CASP) that binds within the periplasmic cleft of an RND transporter protein (MtrD) produced by Neisseria gonorrhoeae. Surprisingly, CASP was able to render rod-shaped Gram-negative bacteria, but not gonococci, susceptible to an antibiotic that is a substrate for the gonococcal MtrCDE efflux pump. CASP (or its future derivatives) could be used as an adjuvant to treat infections for which RND efflux contributes to multidrug resistance.
登录
查看更多内容
影响因子：
--
作者：
de la Fuente-Núñez C;Reffuveille F;Mansour SC;Reckseidler-Zenteno SL;Hernández D;Brackman G;Coenye T;Hancock RE
通讯作者：
Hancock RE
影响因子：
3
作者：
Haney EF;Mansour SC;Hilchie AL;de la Fuente-Núñez C;Hancock RE
通讯作者：
Hancock RE
影响因子：
3.7
作者：
Bolla JR;Su CC;Do SV;Radhakrishnan A;Kumar N;Long F;Chou TH;Delmar JA;Lei HT;Rajashankar KR;Shafer WM;Yu EW
通讯作者：
Yu EW
影响因子：
3
作者：
Cox, DL;Sun, YC;Shafer, WM
通讯作者：
Shafer, WM
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K