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@ARTICLE{Sanwald:877532,
author = {Sanwald, Julia L. and Poschmann, Gereon and Stühler, Kai
and Behrends, Christian and Hoffmann, Silke and Willbold,
Dieter},
title = {{T}he {GABARAP} {C}o-{S}ecretome {I}dentified by
{APEX}2-{GABARAP} {P}roximity {L}abelling of {E}xtracellular
{V}esicles},
journal = {Cells},
volume = {9},
number = {6},
issn = {2073-4409},
address = {Basel},
publisher = {MDPI},
reportid = {FZJ-2020-02268},
pages = {1468 -},
year = {2020},
abstract = {The autophagy-related ATG8 protein GABARAP has not only
been shown to be involved in the cellular self-degradation
process called autophagy but also fulfils functions in
intracellular trafficking processes such as receptor
transport to the plasma membrane. Notably, available mass
spectrometry data suggest that GABARAP is also secreted into
extracellular vesicles (EVs). Here, we confirm this finding
by the immunoblotting of EVs isolated from cell culture
supernatants and human blood serum using specific
anti-GABARAP antibodies. To investigate the mechanism by
which GABARAP is secreted, we applied proximity labelling, a
method for studying the direct environment of a protein of
interest in a confined cellular compartment. By expressing
an engineered peroxidase (APEX2)-tagged variant of
GABARAP—which, like endogenous GABARAP, was present in EVs
prepared from HEK293 cells—we demonstrate the
applicability of APEX2-based proximity labelling to EVs. The
biotinylated protein pool which contains the APEX2-GABARAP
co-secretome contained not only known GABARAP interaction
partners but also proteins that were found in
APEX2-GABARAP’s proximity inside of autophagosomes in an
independent study. All in all, we not only introduce a
versatile tool for co-secretome analysis in general but also
uncover the first details about autophagy-based pathways as
possible biogenesis mechanisms of GABARAP-containing EVs.},
cin = {IBI-7},
ddc = {570},
cid = {I:(DE-Juel1)IBI-7-20200312},
pnm = {552 - Engineering Cell Function (POF3-552)},
pid = {G:(DE-HGF)POF3-552},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:32560054},
UT = {WOS:000553607900001},
doi = {10.3390/cells9061468},
url = {https://juser.fz-juelich.de/record/877532},
}