Proteolysis enhanced proximity proteomics
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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 in membrane-protected compartments can be readily enriched and identified.
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Created: 9th Jul 2024 at 13:39
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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
Studies: ACSL3/GABARAPL2 interactor, ADAM10-Mediated Ectodomain Shedding Is an Essential Driver of Podocyte D..., AMPK/Lysosomal Damage/Galectins, An optimized quantitative proteomics method establishes the cell type-re..., Autophagosome content profiling, Autophagy cargo/CD4+ T cell proliferation, BAG3-mediated autophagy, Basic Fibroblast Growth Factor 2-Induced Proteome Changes Endorse Lewy B..., Beneficial Effect of ACI-24 Vaccination on Aβ Plaque Pathology and Micro..., C9orf72/UBR5/Quality control, CCNF/ALS, CNN2/Lysosome damage, COP9 signalosome/Ischemic stroke, Cell-type-specific profiling of brain mitochondria reveals functional an..., Deciphering sources of PET signals in the tumor microenvironment of glio..., Endothelial Aging, Experimental evidence for temporal uncoupling of brain Aβ deposition and..., Fibrillar Aβ triggers microglial proteome alterations and dysfunction in..., Loss of NPC1 enhances phagocytic uptake and impairs lipid trafficking in..., Lysosomal targeting of TAPL, NDP52 interactome, Neuronal differentiation of LUHMES cells is accompanied by substantial c..., NrCAM is a marker for substrate-selective activation of ADAM10 in Alzhei..., Pro-inflammatory activation following demyelination is required for myel..., Protein content of autophagic vesicles, Proteolysis enhanced proximity proteomics, Proteomic and lipidomic profiling of demyelinating lesions identifies fa..., Proteomic profiling in cerebral amyloid angiopathy reveals an overlap wi..., Proteomics of inflammasome activation for rapid immune responses, Proteomics of mouse brain endothelium uncovers dysregulation of vesicula..., Reactivated endogenous retroviruses promote protein aggregate spreading, Signal peptide peptidase-like 2c impairs vesicular transport and cleaves..., Signatures of glial activity can be detected in the CSF proteome, Spatial proteomics in three-dimensional intact specimens, Spatial proteomics reveals secretory pathway disturbances caused by neur..., TBK1/ALS, TECPR2/secretory pathway, Targeting the TCA cycle can ameliorate widespread axonal energy deficien..., The Alzheimer's disease-linked protease BACE1 modulates neuronal IL-6 si..., The Alzheimer's disease-linked protease BACE2 cleaves VEGFR3 and modulat..., The intramembrane protease SPPL2c promotes male germ cell development by..., The pseudoprotease iRhom1 controls ectodomain shedding of membrane prote..., The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor com..., UBE2QL1/Lysosome damage, UBQLN2/ALS
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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
he 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 ...
Creators: Rainer Malik, Christian Behrends
Submitter: Rainer Malik
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