Fig. 5.

TurboID labeling and streptavidin affinity-purified (AP) captures cellularly distinct proteomes. A, differential expression analysis (DEA) of BV2 AP samples showing robust TurboID biotinylation of over >1700 proteins over endogenously biotinylated proteins derived from untransduced cell lines. B, DEA depicting biotin enrichment of N2A transduced AP proteome reveals >2000 proteins abelled by TurboID. C, DEA comparing transduced AP proteomes of BV2 (left) and N2A (right) TurboID-biotinylated proteomes. There are 936 proteins labeled by TurboID enriched in BV2 and 404 proteins enriched in N2A biotin-labeled proteomes. Proteins with disease relevance to neurodegenerative disease are highlighted. D, schematic of the variety of subcellular compartments abelled by TurboID in microglia (BV2). E, Gene Ontology (GO) of highly enriched cellular components and biological processes within the biotin-labeled BV2 proteome. TurboID biotinylates translational machinery, endosomal machinery, and vesicle-bound membranes in BV2 cells. F, schematic of diversity of subcellular compartments abelled by TurboID in synaptic compartment (N2A). G, GO of significantly enriched cellular components and biological processes within the biotin-labeled N2A proteome confirms that TurboID biotinylates neuronal processes including synaptic machinery and neuron projection.