Autophagosome content profiling - STAR Protocol Data Set Version 1
External Linkhe ascorbate peroxidase APEX2 is commonly used to study the neighborhood of a protein of interest by proximity-dependent biotinylation. Here, we describe a protocol for sample processing compatible with immunoblotting and mass spectrometry that is suitable to specifically map the content of autophagosomes and potentially other transient vesicles without the need of subcellular fractionation. By combining live-cell biotinylation with proteinase K digestion of cell homogenates, proteins enriched in membrane-protected compartments can be readily enriched and identified.
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Created: 9th Jul 2024 at 13:40
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Version 1 (earliest) Created 9th Jul 2024 at 13:40 by Rainer Malik
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Projects: SyNergy - published datasets
Institutions: DZNE
Projects: SyNergy - published datasets
Institutions: Klinikum der Universität München
Public web page: Not specified
Submitter: Rainer Malik
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The ascorbate peroxidase APEX2 is commonly used to study the neighborhood of a protein of interest by proximity-dependent biotinylation. Here, we describe a protocol for sample processing compatible with immunoblotting and mass spectrometry that is suitable to specifically map the content of autophagosomes and potentially other transient vesicles without the need of subcellular fractionation. By combining live-cell biotinylation with proteinase K digestion of cell homogenates, proteins enriched ...
Submitter: Rainer Malik
Investigation: Proteomics
Assays: Affinity purification coupled with mass spectrometry proteomics
Samples were essentially prepared as described in Zellner et al. Mol Cell 2021 (DOI: 10.1016/j.molcel.2021.01.009)
Submitter: Rainer Malik
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Proteomics
Abstract (Expand)
Authors: S. Zellner, K. Nalbach, C. Behrends
Date Published: 18th Jun 2021
Publication Type: Journal
