Biosensor for Multimodal Characterization of an Essential ABC Transporter for Next-Generation Antibiotic Research.
Biosensor for Multimodal Characterization of an Essential ABC Transporter for Next-Generation Antibiotic Research.
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DOI：
10.1021/acsami.2c21556
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发表时间：
2023-03-15
影响因子：
9.5
通讯作者：
van Veen HW
中科院分区：
文献类型：
--
作者：
Bali K;Guffick C;McCoy R;Lu Z;Kaminski CF;Mela I;Owens RM;van Veen HW
随着抗生素耐药性威胁的增加，开发针对多药耐药性尤其根深蒂固和令人担忧的致病菌的抗菌剂成为一个特别的重点。新型抗菌剂的一个这样的靶标是atp结合盒（ABC）转运体MsbA，它存在于革兰氏阴性致病菌的质膜中，对这些细菌的生存至关重要。负载型脂质双分子层（slb）可以与多种光学、生化和电化学技术相结合，在监测膜蛋白结构和功能方面非常有用。在这里，我们形成含有大肠杆菌MsbA的slb，并使用原子力显微镜（AFM）和结构照明显微镜（SIM）作为高分辨率显微镜技术来研究slb和掺入的MsbA蛋白的完整性。然后，我们将这些slb集成到基于导电聚合物聚（3,4-乙烯二氧基噻吩）聚苯乙烯磺酸盐（PEDOT:PSS）的微电极阵列（MEA）上，利用电化学阻抗谱（EIS）监测ATP水解反应中MsbA蛋白的离子流动。这些EIS测量可以与msba - atp酶活性的生化检测相关联。为了证明这种SLB方法的潜力，我们不仅观察了野生型MsbA的活性，还观察了两个先前表征的突变体以及喹啉类MsbA抑制剂G907的活性，以表明EIS系统可以检测ABC转运蛋白活性的变化。我们的工作结合了多种技术来彻底研究脂质双分子层中的MsbA以及这种蛋白质的潜在抑制剂的作用。我们设想这个平台将促进下一代抗菌剂的开发，抑制微生物中的MsbA或其他必需的膜转运蛋白。
As the threat of antibiotic resistance increases, there is a particular focus on developing antimicrobials against pathogenic bacteria whose multidrug resistance is especially entrenched and concerning. One such target for novel antimicrobials is the ATP-binding cassette (ABC) transporter MsbA that is present in the plasma membrane of Gram-negative pathogenic bacteria where it is fundamental to the survival of these bacteria. Supported lipid bilayers (SLBs) are useful in monitoring membrane protein structure and function since they can be integrated with a variety of optical, biochemical, and electrochemical techniques. Here, we form SLBs containing Escherichia coli MsbA and use atomic force microscopy (AFM) and structured illumination microscopy (SIM) as high-resolution microscopy techniques to study the integrity of the SLBs and incorporated MsbA proteins. We then integrate these SLBs on microelectrode arrays (MEA) based on the conducting polymer poly(3,4-ethylenedioxy-thiophene) poly(styrene sulfonate) (PEDOT:PSS) using electrochemical impedance spectroscopy (EIS) to monitor ion flow through MsbA proteins in response to ATP hydrolysis. These EIS measurements can be correlated with the biochemical detection of MsbA-ATPase activity. To show the potential of this SLB approach, we observe not only the activity of wild-type MsbA but also the activity of two previously characterized mutants along with quinoline-based MsbA inhibitor G907 to show that EIS systems can detect changes in ABC transporter activity. Our work combines a multitude of techniques to thoroughly investigate MsbA in lipid bilayers as well as the effects of potential inhibitors of this protein. We envisage that this platform will facilitate the development of next-generation antimicrobials that inhibit MsbA or other essential membrane transporters in microorganisms.
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影响因子：
5.9
作者：
Guo D;Singh H;Shimoyama A;Guffick C;Tang Y;Rowe SM;Noel T;Spring DR;Fukase K;van Veen HW
通讯作者：
van Veen HW
来自海平球菌的一种未表征的主要促进剂超家族转运蛋白具有 Na (Li , K )/H 反向转运蛋白和多药物外排泵的双重功能
影响因子：
5.2
作者：
Abdel-Motaal H;Meng L;Zhang Z;Abdelazez AH;Shao L;Xu T;Meng F;Abozaed S;Zhang R;Jiang J
通讯作者：
Jiang J
影响因子：
4.1
作者：
Eckford, Paul D. W.;Sharom, Frances J.
通讯作者：
Sharom, Frances J.
DOI：
10.1038/nrm2668
发表时间：
2009-05
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.6
作者：
KAROW, M;GEORGOPOULOS, C
通讯作者：
GEORGOPOULOS, C