Proximity-proteomics-based autophagosome content profiling

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Autophagy allows the degradation of cytosolic endogenous and exogenous material in the lysosome. Substrates are engulfed by double-membrane vesicles, coined autophagosomes, which subsequently fuse with lysosomes. Depending on the involvement of specific receptor proteins, autophagy occurs in a selective or nonselective manner. While this process is well understood at the level of bulky cargo such as mitochondria and bacteria, we know very little about individual proteins and protein complexes ...

Proximity-proteomics-based autophagosome content profiling to identify a role for LC3C in maintaining basal mitochondrial homeostasis. Selected mitochondrial proteins, including MTX1, were targeted by LC3C and p62 through a piecemeal mitophagy pathway. SILAC cells biotinylated using APEX proximity labeling. Cell lysates treated with protease. RIPA soluble and insoluble fractions were subjected to Streptavidin pulldown followed by in gel digestion. 4 lanes were cut from one sample.