Cationic Amphipathic Antimicrobial Peptides Perturb the Inner Membrane of Germinated Spores Thus Inhibiting Their Outgrowth.
Cationic Amphipathic Antimicrobial Peptides Perturb the Inner Membrane of Germinated Spores Thus Inhibiting Their Outgrowth.
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DOI：
10.3389/fmicb.2018.02277
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发表时间：
2018
影响因子：
5.2
通讯作者：
Brul S
中科院分区：
文献类型：
--
作者：
Omardien S;Drijfhout JW;Zaat SA;Brul S
研究了四种阳离子两性抗菌肽（AMPs）对非致病性革兰氏阳性芽孢杆菌枯草芽孢杆菌的作用模式。抗菌肽分别为TC19、TC84、BP2和抗生素Nisin A. TC19和TC84来源于人凝血素-1。杀菌肽2 （BP2）来源于人杀菌通透性增加蛋白（BPI）。我们使用结构照明显微镜（SIM）、荧光显微镜、Alexa 488标记的TC84、产生与绿色荧光蛋白（GFP）融合的蛋白质的枯草芽孢杆菌突变体和单细胞活成像来确定肽对孢子的作用。结果表明，TC19、TC84、BP2和Nisin A对萌发的孢子具有抑菌作用，其作用方式是扰乱孢子的内膜，从而阻止孢子长成营养细胞。单细胞活体成像结果显示，AMPs不影响发芽过程，但影响营养细胞的破裂时间和后代时间。Alexa 488标记的TC84表明TC84可能与休眠孢子外壳结合。因此，在单细胞活显像期间，休眠孢子也预先包被amp，并在无amp的培养基上培养。TC19、TC84和BP2对孢子萌发过程无影响，但对萌发时间和产生时间有不同的影响。然而，预涂Nisin A后，孢子破裂并长成营养细胞的比例明显降低，表明Nisin A对孢子具有新的应用潜力。我们的发现有助于理解抗菌肽，并显示抗菌肽作为抗孢子形成细菌的最终治疗剂的潜力。
The mode of action of four cationic amphipathic antimicrobial peptides (AMPs) was evaluated against the non-pathogenic, Gram-positive, spore-forming bacterium, Bacillus subtilis. The AMPs were TC19, TC84, BP2, and the lantibiotic Nisin A. TC19 and TC84 were derived from the human thrombocidin-1. Bactericidal peptide 2 (BP2) was derived from the human bactericidal permeability increasing protein (BPI). We employed structured illumination microscopy (SIM), fluorescence microscopy, Alexa 488-labeled TC84, B. subtilis mutants producing proteins fused to the green fluorescent protein (GFP) and single-cell live imaging to determine the effects of the peptides against spores. TC19, TC84, BP2, and Nisin A showed to be bactericidal against germinated spores by perturbing the inner membrane, thus preventing outgrowth to vegetative cells. Single cell live imaging showed that the AMPs do not affect the germination process, but the burst time and subsequent generation time of vegetative cells. Alexa 488-labeled TC84 suggested that the TC84 might be binding to the dormant spore-coat. Therefore, dormant spores were also pre-coated with the AMPs and cultured on AMP-free culture medium during single-cell live imaging. Pre-coating of the spores with TC19, TC84, and BP2 had no effect on the germination process, and variably affected the burst time and generation time. However, the percentage of spores that burst and grew out into vegetative cells was drastically lower when pre-coated with Nisin A, suggesting a novel application potential of this lantibiotic peptide against spores. Our findings contribute to the understanding of AMPs and show the potential of AMPs as eventual therapeutic agents against spore-forming bacteria.
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影响因子：
4.4
作者：
Abriouel, H;Maqueda, M;Valdivia, E
通讯作者：
Valdivia, E
DOI：
10.1073/pnas.0306859101
发表时间：
2004-05-18
影响因子：
11.1
作者：
Cowan, AE;Olivastro, EM;Setlow, P
通讯作者：
Setlow, P
影响因子：
2.9
作者：
Arnusch, Christopher J.;Branderhorst, Hilbert;Pieters, Roland J.
通讯作者：
Pieters, Roland J.
影响因子：
3.2
作者：
Keijser, Bart J. F.;Ter Beek, Alex;Brul, Stanley
通讯作者：
Brul, Stanley
影响因子：
3.2
作者：
Korza, George;Setlow, Barbara;Setlow, Peter
通讯作者：
Setlow, Peter