Generation of the heterogeneity of extracellular vesicles by membrane organization and sorting machineries
Generation of the heterogeneity of extracellular vesicles by membrane organization and sorting machineries
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DOI：
10.1016/j.bbagen.2019.01.015
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发表时间：
2019-04-01
影响因子：
3
通讯作者：
Maruyama, Ikuro
中科院分区：
文献类型：
--
作者：
Harada, Yoichiro;Suzuki, Takehiro;Maruyama, Ikuro
背景细胞通过未知的机制分泌细胞外囊泡（EV）的异质群体。EV的生物合成已被假定涉及脂质蛋白质簇，也被称为membrane microdomains.Methods：膜的性质和异质性的黑色素瘤衍生的EV进行了分析，通过洗涤剂溶解试验，蔗糖密度梯度超离心和免疫沉淀。EV分泌的RNA干扰和药物treatment.Results调制：我们确定了两个EV膜（低密度外泌体洗涤剂不溶性膜[EV-DIMs]; EV洗涤剂可溶性膜[EV-DSMs]），并发现了一个丰富的，新型的高密度EV-DIMs。高密度EV-DIM积累了微域驻留蛋白flotillin-1，以及包含蛋白10（Adam 10）的去整合素和金属蛋白酶结构域，肝细胞生长因子受体Met及其蛋白水解片段。低密度EV-DIM还含有flotillin-1。EV-DSM富含四跨膜蛋白CD 81、黑素生成酶和Adam 10的蛋白水解片段。不同的EV分泌完整和片段形式的Adam 10，其存在于不同的膜类型中。Met的片段形式与DIM的结合比完整形式有效得多，并且它们由不同的EV分泌。我们确定，运输机械所需的内体分选复合物是必不可少的EV分泌的成熟和碎片形式的Adam 10和Met.Conclusion：本研究的结果揭示了膜组织和分选机械之间的相互作用产生的EVs.General意义的异质性的作用：这项研究提供了新的见解EV生物起源的重要方面。
Background Cells secrete heterogeneous populations of extracellular vesicles (EVs) via unknown mechanisms. EV biogenesis has been postulated to involve lipid-protein clusters, also known as membrane microdomains.Methods: Membrane properties and heterogeneity of melanoma-derived EVs were analyzed by a detergent solubilization assay, sucrose density gradient ultracentrifugation and immunoprecipitation. EV secretion was modulated by RNA interference and pharmacological treatments.Results: We identified two EV membranes (low-density exosomal detergent-insoluble membranes [EV-DIMs]; EV detergent-soluble membranes [EV-DSMs]) and discovered an abundant, novel type of high-density EV-DIMs. The high-density EV-DIMs accumulated the microdomain-resident protein flotillin-1, as well as a disintegrin and metalloproteinase domain containing protein 10 (Adam10), the hepatocyte growth factor receptor Met and its proteolytic fragments. Low-density EV-DIMs also contained flotillin-1. EV-DSMs were enriched with tetraspanin CD81, melanogenic enzymes and proteolytic fragments of Adam10. Intact and fragmented forms of Adam10, which resided in distinct membrane types, were secreted by different EVs. The fragmented form of Met was associated with DIMs much more efficiently than the intact form and they were secreted by distinct EVs. We identified that the endosomal sorting complexes required for transport machinery was indispensable for EV secretion of both mature and fragmented forms of Adam10 and Met.Conclusion: The findings of this study reveal the role of the interplay between membrane organization and sorting machineries in generating the heterogeneity of EVs.General significance: This study provides novel insights into important aspects of EV biogenesis.