2019
DOI: 10.1021/acsinfecdis.9b00299
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Fluorescence Imaging of Bacterial Killing by Antimicrobial Peptide Dendrimer G3KL

Abstract: We recently discovered that peptide dendrimers such as G3KL ((KL)8(KKL)4(KKL)2 KKL, K = branching l-lysine) exert strong activity against Gram-negative bacteria including Pseudomonas aeruginosa, Acinetobacter baumannii, and Escherichia coli. Herein, we report a detailed mechanistic study using fluorescence labeled analogs bearing fluorescein (G3KL-Fluo) or dansyl (G3KL-Dansyl), which show a similar bioactivity profile as G3KL. Imaging bacterial killing by super-resolution stimulated emission depletion (STED) m… Show more

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Cited by 61 publications
(73 citation statements)
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“…While theoretically, the resolution in STED is not limited, practically considering the phototoxicity tolerance of the biological sample and the laser power available, an axial resolution of ~35 nm and laterally of ~90 nm can be achieved [ 38 ]. With the advent of easier sample preparation and the availability of commercial STED microscopes, STED microscopy is an emerging option for investigating questions in biomedical research, including host–pathogen and AMP–pathogen interactions [ 39 , 40 , 41 ]. In this study, we evaluated the suitability of innovative STED microscopy as a tool for directly observing host–pathogen interaction using, as an example, LL-37, primary human macrophages and virulent Mtb .…”
Section: Introductionmentioning
confidence: 99%
“…While theoretically, the resolution in STED is not limited, practically considering the phototoxicity tolerance of the biological sample and the laser power available, an axial resolution of ~35 nm and laterally of ~90 nm can be achieved [ 38 ]. With the advent of easier sample preparation and the availability of commercial STED microscopes, STED microscopy is an emerging option for investigating questions in biomedical research, including host–pathogen and AMP–pathogen interactions [ 39 , 40 , 41 ]. In this study, we evaluated the suitability of innovative STED microscopy as a tool for directly observing host–pathogen interaction using, as an example, LL-37, primary human macrophages and virulent Mtb .…”
Section: Introductionmentioning
confidence: 99%
“…G3KL is a representative of these third-generation dendrimers, which shows activity against MDR Pseudomonas aeruginosa clinical isolates as well as Acinetobacter baumannii and Enterobacteriaceae harboring extended-spectrum ␤-lactamases (ESBLs) (21,22). G3KL has low hemolytic activity, with minimal hemolytic concentrations above 2,000 g/ml (21) and IC 50 values around 1,000 g/ml for HeLa and CHO cells (23). G3KL and its derivatives are synthesized with D-amino acids conferring increased stability against proteolytic cleavage in the presence of human serum (21).…”
mentioning
confidence: 99%
“…G3KL AMPD showed as well to inhibit P. aeruginosa biofilm formation [100]. Fluorescently labeled G3KL entirely diffused into P. aeruginosa within 15 min as shown in a time-lapse assay, suggesting AMPD diffusion or translocation into the cytoplasm through damaging both the outer and inner bacterial membrane, as was confirmed by TEM [101]. In addition to their intrinsic activity and due to the fact that AMPDs have different functional groups, it makes them very attractive delivery systems as drug carriers.…”
Section: Antimicrobial Dendrimer Peptidesmentioning
confidence: 69%