Lipid and protein content profiling of isolated native autophagic vesicles

dc.contributor.authorSchmitt, Daniel
dc.contributor.authorBozkurt, Süleyman
dc.contributor.authorHenning-Domres, Pascale
dc.contributor.authorHuesmann, Heike
dc.contributor.authorEimer, Stefan
dc.contributor.authorBindila, Laura
dc.contributor.authorBehrends, Christian
dc.contributor.authorBoyle, Emily
dc.contributor.authorWilfling, Florian
dc.contributor.authorTascher, Georg
dc.contributor.authorMünch, Christian
dc.contributor.authorBehl, Christian
dc.contributor.authorKern, Andreas
dc.date.accessioned2023-01-10T10:27:27Z
dc.date.available2023-01-10T10:27:27Z
dc.date.issued2022
dc.description.abstractAutophagy is responsible for clearance of an extensive portfolio of cargoes, which are sequestered into vesicles, called autophagosomes, and are delivered to lysosomes for degradation. The pathway is highly dynamic and responsive to several stress conditions. However, the phospholipid composition and protein contents of human autophagosomes under changing autophagy rates are elusive so far. Here, we introduce an antibody-based FACS-mediated approach for the isolation of native autophagic vesicles and ensured the quality of the preparations. Employing quantitative lipidomics, we analyze phospholipids present within human autophagic vesicles purified upon basal autophagy, starvation, and proteasome inhibition. Importantly, besides phosphoglycerides, we identify sphingomyelin within autophagic vesicles and show that the phospholipid composition is unaffected by the different conditions. Employing quantitative proteomics, we obtain cargo profiles of autophagic vesicles isolated upon the different treatment paradigms. Interestingly, starvation shows only subtle effects, while proteasome inhibition results in the enhanced presence of ubiquitin–proteasome pathway factors within autophagic vesicles. Thus, here we present a powerful method for the isolation of native autophagic vesicles, which enabled profound phospholipid and cargo analyses.en_GB
dc.description.sponsorshipGefördert durch die Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 491381577de
dc.identifier.doihttp://doi.org/10.25358/openscience-8543
dc.identifier.urihttps://openscience.ub.uni-mainz.de/handle/20.500.12030/8559
dc.language.isoengde
dc.rightsCC-BY-NC-ND-4.0*
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subject.ddc610 Medizinde_DE
dc.subject.ddc610 Medical sciencesen_GB
dc.titleLipid and protein content profiling of isolated native autophagic vesiclesen_GB
dc.typeZeitschriftenaufsatzde
jgu.journal.titleEMBO reportsde
jgu.journal.volume23de
jgu.organisation.departmentFB 04 Medizinde
jgu.organisation.nameJohannes Gutenberg-Universität Mainz
jgu.organisation.number2700
jgu.organisation.placeMainz
jgu.organisation.rorhttps://ror.org/023b0x485
jgu.pages.alternativee53065de
jgu.publisher.doi10.15252/embr.202153065de
jgu.publisher.issn1469-3178de
jgu.publisher.nameEMBO Pressde
jgu.publisher.placeHeidelbergde
jgu.publisher.year2022
jgu.rights.accessrightsopenAccess
jgu.subject.ddccode610de
jgu.subject.dfgLebenswissenschaftende
jgu.type.dinitypeArticleen_GB
jgu.type.resourceTextde
jgu.type.versionPublished versionde

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