Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Medical University of Gdansk, 107 Hallera Street, 80-416 Gdansk, Poland.
Department of Physical Chemistry, Faculty of Pharmacy, Medical University of Gdansk, 107 Hallera Street, 80-416 Gdansk, Poland.
J Chromatogr A. 2020 Jun 21;1621:461047. doi: 10.1016/j.chroma.2020.461047. Epub 2020 Mar 13.
The extracellular vesicles (EVs) released by plant pathogens of the Pectobacterium genus were investigated. The isolates were obtained using differential centrifugation followed by filtration and were characterized in terms of total protein content and particle size distribution. The transmission electron microscopy (TEM) analysis revealed the presence of two morphologically differentiated subpopulations of vesicles in the obtained isolates. The proteomic analysis using matrix-assisted laser desorption ionization mass spectrometry with time of flight detector (MALDI-TOF/TOF-MS) enabled to identify 62 proteomic markers commonly found in EVs of Gram-negative rods from the Enterobacteriaceae family. Capillary electrophoresis (CE) was proposed as a novel tool for the characterization of EVs. The method allowed for automated and fast (<15 min per sample) separation of vesicles from macromolecular aggregates with low sample consumption (about 10 nL per analysis). The approach required simple background electrolyte (BGE) composed of 50 mM BTP and 75 mM glycine (pH 9.5) and standard UV detection. The report presents a new opportunity for quality control of samples containing EVs.
研究了果胶杆菌属植物病原菌释放的细胞外囊泡（EVs）。使用差速离心、过滤分离获得分离株，并从总蛋白含量和粒径分布两方面对其进行了表征。透射电子显微镜（TEM）分析显示，所获得的分离株中存在两种形态上有区别的囊泡亚群。基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/TOF-MS）的蛋白质组学分析能够鉴定出肠杆菌科革兰氏阴性菌 EVs 中常见的 62 种蛋白质组学标记物。毛细管电泳（CE）被提议作为 EVs 表征的新工具。该方法允许在低样品消耗（每次分析约 10 nL）的情况下，自动且快速（<15 分钟/样品）分离囊泡和大分子聚集体。该方法仅需要由 50 mM BTP 和 75 mM 甘氨酸（pH 9.5）组成的简单背景电解质（BGE）和标准 UV 检测。该报告为含有 EVs 的样品的质量控制提供了新的机会。
