Extracellular Vesicles Containing Ceramide-Rich Platforms: "Mobile Raft" Isolation and Analysis
Extracellular Vesicles Containing Ceramide-Rich Platforms: "Mobile Raft" Isolation and Analysis
复制标题
DOI：
10.1007/978-1-0716-0814-2_5
复制
发表时间：
2021-01-01
期刊：
影响因子：
--
通讯作者：
Bieberich, Erhard
中科院分区：
文献类型：
--
作者：
Elsherbini, Ahmed;Qin, Haiyan;Bieberich, Erhard
细胞外囊泡（EV）由真核细胞分泌并充当多种细胞信号传导因子（包括RNA、蛋白质和脂质）的载体。我们描述了一种独特的EV群体，其膜高度富含鞘脂神经酰胺。我们认为，神经酰胺在EV膜组织在富含神经酰胺的平台（CRP），一种类型的脂筏介导的相互作用的神经酰胺与神经酰胺相关蛋白（CAPs）。在这里，我们描述了使用抗神经酰胺抗体分离富含神经酰胺的EV并在摄取到受体细胞中后检测外泌体的方法。此外，我们还讨论了使用纳米粒子跟踪和质谱法进行EV分析的方法。所述方法可以扩展到使用针对这些EV特异性的脂质的抗体分离其他类型的EV和由EV从供体细胞转运到受体细胞的“移动的筏”。
Extracellular vesicles (EVs) are secreted by eukaryotic cells and serve as carriers for a variety of cell signaling factors, including RNAs, proteins, and lipids. We described a unique population of EVs, the membrane of which is highly enriched with the sphingolipid ceramide. We suggested that ceramide in the EV membrane is organized in ceramide-rich platforms (CRPs), a type of lipid raft that mediates interaction of ceramide with ceramide-associated proteins (CAPs). Here, we describe methods using anti-ceramide antibody to isolate ceramide-enriched EVs and detect exosomes after uptake into recipient cells. In addition, we discuss methods for EV analysis using nanoparticle tracking and mass spectrometry. The methods can be extended to the isolation of other types of EVs and "mobile rafts" transported by EVs from donor to recipient cells using antibodies against lipids specific for these EVs.