Arrestin-Domain Containing Protein 1 (Arrdc1) Regulates the Protein Cargo and Release of Extracellular Vesicles
Arrestin-Domain Containing Protein 1 (Arrdc1) Regulates the Protein Cargo and Release of Extracellular Vesicles
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DOI：
10.1002/pmic.201800266
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发表时间：
2018-09-01
期刊：
影响因子：
3.4
通讯作者：
Kumar, Sharad
中科院分区：
文献类型：
--
作者：
Anand, Sushma;Foot, Natalie;Kumar, Sharad
细胞外囊泡（EV）是由多种细胞类型释放的含有核酸和蛋白质的脂质双层囊泡。人们对货物如何包装到电动汽车中知之甚少。蛋白质的泛素化是一种关键的翻译后修饰，其调节蛋白质稳定性和向亚细胞区室（包括EV）的运输。最近，含有抑制蛋白结构域的蛋白1（Arrdc 1）是遍在蛋白连接酶Nedd 4家族的衔接子，它与外体（EV的一种亚型，从质膜上发芽）的释放有关。然而，目前尚不清楚Arrdc 1是否可以调节外泌体的释放，外泌体是一类内吞衍生的EV。此外，目前还不清楚Arrdc 1是否可以调节蛋白质货物进入电动汽车的分选。外泌体和外泌体分离自小鼠胚胎成纤维细胞，所述小鼠胚胎成纤维细胞分离自野生型和Arrdc 1缺陷型（Arrdc 1（-/-））小鼠。基于纳米粒子跟踪分析的EV定量表明，Arrdc 1调节外泌体和外泌体的释放。蛋白质组学分析突出了Arrdc 1缺失后EV中蛋白质货物的变化。功能富集分析揭示了线粒体蛋白质在外泌体中的富集，而与细胞粘附蛋白的凋亡裂解和皮质包膜的形成有关的蛋白质在Arrdc 1敲除后在外泌体中显著耗尽。
Extracellular vesicles (EVs) are lipid-bilayered vesicles that are released by multiple cell types and contain nucleic acids and proteins. Very little is known about how the cargo is packaged into EVs. Ubiquitination of proteins is a key posttranslational modification that regulates protein stability and trafficking to subcellular compartments including EVs. Recently, arrestin-domain containing protein 1 (Arrdc1), an adaptor for the Nedd4 family of ubiquitin ligases, has been implicated in the release of ectosomes, a subtype of EV that buds from the plasma membrane. However, it is currently unknown whether Arrdc1 can regulate the release of exosomes, a class of EVs that are derived endocytically. Furthermore, it is unclear whether Arrdc1 can regulate the sorting of protein cargo into the EVs. Exosomes and ectosomes are isolated from mouse embryonic fibroblasts isolated from wild type and Arrdc1-deficient (Arrdc1(-/-)) mice. Nanoparticle tracking analysis-based EV quantitation shows that Arrdc1 regulates the release of both exosomes and ectosomes. Proteomic analysis highlights the change in protein cargo in EVs upon deletion of Arrdc1. Functional enrichment analysis reveals the enrichment of mitochondrial proteins in ectosomes, while proteins implicated in apoptotic cleavage of cell adhesion proteins and formation of cornified envelope are significantly depleted in exosomes upon knockout of Arrdc1.