PXD040569
PXD040569 is an original dataset announced via ProteomeXchange.
Dataset Summary
Title | TurboID-EV: a proximity-labeling proteomic approach to dissect the uptake mechanism of extracellular vesicles |
Description | Extracellular vesicles (EVs), including exosomes, have been recognized as key mediators of intercellular communications through donor EV and recipient cell interaction. Until now, most studies have focused on the development of analytical tools to separate EVs and their applications for molecular profiling of EV cargo. However, we lack a complete picture of the mechanism of uptake of EVs by recipient cells. Here, we developed the TurboID-EV system with the engineered biotin ligase, TurboID, tethered to the EV membrane, which allowed us to track the footprints of EVs during and after EV uptake by proximity-dependent biotinylation of recipient cellular proteins. To enrich and analyze biotinylated recipient proteins from low amounts of input cells (corresponding to ~10 µg proteins), we developed an integrated proteomic workflow that combined SILAC labeling, fluorescence-activated cell sorting, spintip-based streptavidin affinity purification, and mass spectrometry. Using the method, we successfully identified 456 biotinylated recipient proteins, including not only well-known proteins involved in endocytosis and macropinocytosis but also previously unappreciated membrane-associated proteins such as DSP and JUP. The TurboID-EV system should be readily applicable to various EV subtypes and recipient cell types, providing a promising tool to dissect the specificity of EV uptake mechanisms on a proteome-wide scale. |
HostingRepository | jPOST |
AnnounceDate | 2023-08-21 |
AnnouncementXML | Submission_2023-08-20_08:00:04.977.xml |
DigitalObjectIdentifier | |
ReviewLevel | Non peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Yuka Li |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | S-carboxamidomethyl-L-cysteine; alpha-amino acetylated residue; L-methionine sulfoxide |
Instrument | Orbitrap Fusion Lumos; Orbitrap Exploris 480 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
---|---|---|---|
0 | 2023-03-03 05:16:42 | ID requested | |
⏵ 1 | 2023-08-20 08:00:05 | announced |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: Extracellular vesicle, Proximity labeling |
Contact List
Yasushi Ishihama | |
---|---|
lab head | |
Yuka Li | |
contact affiliation | Kyoto University |
dataset submitter |
Full Dataset Link List
jPOST dataset URI |
Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.jpostdb.org/JPST002070/ |