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We describe a tool, Spatio-Temporal Association Mapping of Proteins (STAMP), for identifying protein interactomes via proximity labeling. For a proof-of-principle study, we use cytidine 5'-triphosphate synthase (CTPS) as an example. CTPS, a metabolic enzyme, forms filamentous structures termed cytoophidia in various tissues. We apply STAMP to a variety of developmental stages and tissues in Drosophila including adult ovaries. Using a cell-specific GAL4 driver, we verify that TurboID can biotinylate the bait protein CTPS, making possible the identification of protein-protein interactions (PPIs) in individual cells. Using the wild-type and mutant CTPS as bait proteins, STAMP results in two distinct sets of proximate proteomes. Our results suggest that STAMP is a feasible tool to catch in vivo PPIs in situ at a defined spatiotemporal resolution.

Original publication

DOI

10.1007/978-1-0716-2970-3_20

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2023

Volume

2626

Pages

365 - 379

Keywords

Drosophila, Ovary, Proximity labeling, STAMP, TurboID, Animals, Female, Carbon-Nitrogen Ligases, Cytoskeleton, Drosophila, Ovary, Proteome