UBL3 modification influences protein sorting to small extracellular vesicles
Hiroshi AgetaNatsumi Ageta‐IshiharaKeisuke HitachiÖzge KarayelTakanori OnouchiHisateru YamaguchiTomoaki KahyoKen HatanakaKoji IkegamiYusuke YoshiokaKenji NakamuraNobuyoshi KosakaMasashi NakataniAkiyoshi UezumiTomihiko IdeYutaka TsutsumiHaruhiko SugimuraMakoto KinoshitaTakahiro OchiyaMatthias MannMitsutoshi SetouKunihiro Tsuchida
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Abstract Exosomes, a type of small extracellular vesicles (sEVs), derived from multivesicular bodies (MVBs), mediate cell-to-cell communication by transporting proteins, mRNAs, and miRNAs. However, the molecular mechanism by which proteins are sorted to sEVs is not fully understood. Here, we report that ubiquitin-like 3 (UBL3)/membrane-anchored Ub-fold protein (MUB) acts as a posttranslational modification (PTM) factor that regulates protein sorting to sEVs. We find that UBL3 modification is indispensable for sorting of UBL3 to MVBs and sEVs. We also observe a 60% reduction of total protein levels in sEVs purified from Ubl3 -knockout mice compared with those from wild-type mice. By performing proteomics analysis, we find 1241 UBL3-interacting proteins, including Ras. We also show that UBL3 directly modifies Ras and oncogenic RasG12V mutant, and that UBL3 expression enhances sorting of RasG12V to sEVs via UBL3 modification. Collectively, these results indicate that PTM by UBL3 influences the sorting of proteins to sEVs.Keywords:
Extracellular Vesicles
Proteome
Cell Sorting
Extracellular vesicles (EVs) are cell-derived lipid vesicles in a size range of 20–1000 nm; often, these are classified as smaller and larger EVs in the literature or also commonly called small EVs (“exosomes”) and medium/large EVs (“microvesicles”) [...]
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(2012). The launch of Journal of Extracellular Vesicles (JEV), the official journal of the International Society for Extracellular Vesicles – about microvesicles, exosomes, ectosomes and other extracellular vesicles. Journal of Extracellular Vesicles: Vol. 1, No. 1, 18514.
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Extracellular vesicles (EVs) are lipid bilayer-bound vesicles secreted by cells. Subtypes of EVs such as microvesicles and exosomes are further categorized mainly by their different biogenesis mechanisms. EVs have been revealed to play an important role in disease diagnosis and intercellular communication. Despite the wide interest in EVs, the technologies for the purification and enrichment of EVs are still in their infancy. The isolation of EVs, especially exosomes, is inherently challenging due to their small size and heterogeneity. In this review, we mainly introduce the advances of techniques in isolating microvesicles and exosomes according to their approaches. Also, we discuss the limitations of currently reported technologies in terms of their specificity and efficiency, and provide our thoughts about future developments of EV purification and enrichment technology.
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