Murgoci Adriana-Natalia, Cardon Tristan, Aboulouard Soulaimane, Duhamel Marie, Fournier Isabelle, Cizkova Dasa, Salzet Michel
Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France; Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava 84510, Slovakia.
Univ. Lille, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse-PRISM, Lille 59000, France.
iScience. 2020 May 22;23(5):101045. doi: 10.1016/j.isci.2020.101045. Epub 2020 Apr 10.
Extracellular vesicles (EVs) mediate intercellular communication and regulate a broad range of biological processes. Novel therapeutic strategies have emerged based on the use of EVs as biological nanoparticles. To separate isolated EVs from protein aggregates and the external part of EVs membrane proteins, we performed a Trypsin/Lys C digestion treatment of EVs pellets, followed by Amicon filtration. After these steps, all the fractions have been subjected to proteomic analyses. Comparison between 6 h Trypsin/Lys C treatment or non-treated EVs revealed a quantitative variation of the surface proteins. Some surface proteins have been demasked after 6 h enzymatic digestion like CD81, CD82, Ust, Vcan, Lamp 1, Rab43, Annexin A2, Synthenin, and VSP37b. Moreover, six ghost proteins have also been identified and one corresponds to a long noncoding RNA. We thus demonstrate the presence of ghost proteins in EVs produced by glioma cells that can contribute to tumorigenesis.
细胞外囊泡（EVs）介导细胞间通讯并调节广泛的生物过程。基于将EVs用作生物纳米颗粒，已经出现了新的治疗策略。为了从蛋白质聚集体和EVs膜蛋白的外部部分分离出分离的EVs，我们对EVs沉淀进行了胰蛋白酶/赖氨酰内切酶C消化处理，然后进行Amicon过滤。经过这些步骤后，所有级分都进行了蛋白质组学分析。6小时胰蛋白酶/赖氨酰内切酶C处理的EVs与未处理的EVs之间的比较揭示了表面蛋白的定量变化。一些表面蛋白在6小时酶消化后被暴露，如CD81、CD82、Ust、Vcan、Lamp 1、Rab43、膜联蛋白A2、合成蛋白和VSP37b。此外，还鉴定出六种幽灵蛋白，其中一种对应于一种长链非编码RNA。因此，我们证明了胶质瘤细胞产生的EVs中存在幽灵蛋白，它们可能有助于肿瘤发生。
