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首页> 外文期刊>Biochemistry >Study of the Multidrug Membrane Transporter of Single Living Pseudomonas aeruginosaCells Using Size-Dependent Plasmonic Nanoparticle Optical Probest
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Study of the Multidrug Membrane Transporter of Single Living Pseudomonas aeruginosaCells Using Size-Dependent Plasmonic Nanoparticle Optical Probest

机译：大小依赖的等离子纳米粒子光学探针研究单活铜绿假单胞菌细胞的多药膜转运蛋白。

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Multidrug membrane transporters (efflux pumps) in both prokaryotes and eukaryotes are res-ponsible for impossible treatments of a wide variety of diseases, including infections and cancer, underscoring the importance of better understanding of their structures and functions for the design of effective therapies. In this study, we designed and synthesized two silver nanoparticles (Ag NPs) with average diameters of 13.1 ± 2.5 nm (8.1-38.6 nm) and 91.0 ± 9.3 nm (56-120 nm) and used the size-dependent plasmonic spectra of single NPs to probe the size-dependent transport kinetics of MexAB-OprM (multidrug transporter) in Pseudomonas aeruginosa in real time at nanometer resolution. We found that the level of accumulation of intracellular NPs in wild-type (WT) cells was higher than in nalB1 (overexpression of MexAB-OprM) but lower than in ΔABM (deletion of MexAB-OprM). In the presence of proton ionophores (CCCP, inhibitor of proton motive force), we found that intracellular NPs in nalB1 were nearly doubled. These results suggest that MexAB-OprM is responsible for the extrusion of NPs out of cells and NPs (orders of magnitude larger than conventional antibiotics) are the substrates of the transporter, which indicates that the substrates may trigger the assembly of the efflux pump optimized for the extrusion of the encountered substrates. We found that the smaller NPs stayed inside the cells longer than larger NPs, suggesting the size-dependent efflux kinetics of the cells. This study shows that multisized NPs can be used to mimic various sizes of antibiotics for probing the size-dependent efflux kinetics of multidrug membrane transporters in single living cells.
机译：原核生物和真核生物中的多药膜转运蛋白（外排泵）负责对包括感染和癌症在内的多种疾病的不可能治疗，从而强调了更好地了解其结构和功能对于设计有效疗法的重要性。在这项研究中，我们设计和合成了两种银纳米颗粒（Ag NPs），它们的平均直径分别为13.1±2.5 nm（8.1-38.6 nm）和91.0±9.3 nm（56-120 nm），并使用了尺寸依赖性的单晶等离子体光谱纳米颗粒以纳米分辨率实时探测铜绿假单胞菌中MexAB-OprM（多药转运蛋白）的大小依赖性转运动力学。我们发现野生型（WT）细胞中细胞内NP的积累水平高于nalB1（MexAB-OprM的过表达），但低于ΔABM（MexAB-OprM的缺失）。在质子离子载体（CCCP，质子动力抑制剂）的存在下，我们发现nalB1中的细胞内NP几乎增加了一倍。这些结果表明，MexAB-OprM负责将NP从细胞中挤出，并且NP（比常规抗生素大几个数量级）是转运蛋白的底物，这表明底物可能会触发为进行优化的外排泵的组装遇到的基材的挤出。我们发现，较小的NP比较大的NP停留在细胞中的时间更长，表明细胞的尺寸依赖性外排动力学。这项研究表明，多种大小的NP可用于模拟各种大小的抗生素，以探测单个活细胞中多种药物膜转运蛋白的大小依赖性外排动力学。

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