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一种互补的 LC-ESI-MS 和 MALDI-TOF 方法,用于筛选养殖欧洲鲈鱼黏液中的抗菌蛋白质组学特征。

A complementary LC-ESI-MS and MALDI-TOF approach for screening antibacterial proteomic signature of farmed European sea bass mucus.

机构信息

Food Technology laboratory, National Institute of Nutrition and Food Technology, Bab Saadoun, 1007 Tunis, Tunisia.

出版信息

Fish Shellfish Immunol. 2013 Aug;35(2):207-12. doi: 10.1016/j.fsi.2013.04.017. Epub 2013 May 2.

DOI:10.1016/j.fsi.2013.04.017
PMID:23643872
Abstract

Antibacterial protection in the mucus is provided by antimicrobial compounds and till now few numbers of AMP and proteins were identified. Herein, mass spectral profiling of fresh mucus from farmed sea bass (Dicentrarchus labrax) using Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometer (MALDI-TOF) and liquid chromatography mass spectrometry is investigated in order to survey the infective/healthy status of the mucus. We identify AMP peptides of 2891.7, 2919.45 and 2286.6 Da molecular weight respectively and characterize Chrysophsins in the mucus of Dicentrarchus labrax. These peptides display broad-spectrum bactericidal activity against Gram-negative (Minimum Inhibitory Concentrations namely MICs < 0.5 μM) and Gram-positive bacteria (MICs < 0.5 μM) including Escherichia coli and Bacillus subtilis. Furthermore, sensitivity to yeast Candida albicans is reported for the first time and shows interesting MICs of less than 2 μM. We also demonstrate that the fish pathogen Aeromonas salmonoicida is sensitive to Chrysophsins (MICs ranging between 5 and 14 μM). Our mucus molecular mass mapping developed approach allows for fast exploration of immune status. Our data provides evidence that Chrysophsins are secreted by immune cells and are released in mucus of non-challenged farmed European sea bass. These results suggest that Chrysophsins, secreted by gills of red sea bream, are an important widespread component of Teleostei defense against disease.

摘要

抗菌保护在黏液中是由抗菌化合物提供的,到目前为止,已经鉴定出少数 AMP 和蛋白质。在此,使用基质辅助激光解吸/电离-飞行时间质谱仪 (MALDI-TOF) 和液相色谱质谱法对养殖鲈鱼(Dicentrarchus labrax)新鲜黏液进行了质谱分析,以调查黏液的感染/健康状况。我们分别鉴定出分子量为 2891.7、2919.45 和 2286.6 Da 的 AMP 肽,并在鲈鱼黏液中表征 Chrysophsins。这些肽对革兰氏阴性(最小抑菌浓度即 MICs <0.5 μM)和革兰氏阳性菌(MICs <0.5 μM)具有广谱杀菌活性,包括大肠杆菌和枯草芽孢杆菌。此外,首次报道了这些肽对酵母白色念珠菌的敏感性,其 MIC 值小于 2 μM。我们还证明,鱼类病原体嗜水气单胞菌对 Chrysophsins 敏感(MIC 值在 5 到 14 μM 之间)。我们开发的黏液分子质量图谱方法允许快速探索免疫状态。我们的数据提供了证据表明,Chrysophsins 是由免疫细胞分泌的,并释放到未受挑战的养殖欧洲鲈鱼的黏液中。这些结果表明,Chrysophsins 是由红鲷鱼的鳃分泌的,是硬骨鱼防御疾病的重要广泛组成部分。

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