Efficient proximity labeling in living cells and organisms with TurboID.

Branon, Tess C; Bosch, Justin A; Sanchez, Ariana D; Udeshi, Namrata D; Svinkina, Tanya; Carr, Steven A; Feldman, Jessica L; Perrimon, Norbert et al. · Nat Biotechnol · 2018

basic_science · Level V

Where this comes from

Abstract

Protein interaction networks and protein compartmentalization underlie all signaling and regulatory processes in cells. Enzyme-catalyzed proximity labeling (PL) has emerged as a new approach to study the spatial and interaction characteristics of proteins in living cells. However, current PL methods require over 18 h of labeling time or utilize chemicals with limited cell permeability or high toxicity. We used yeast display-based directed evolution to engineer two promiscuous mutants of biotin ligase, TurboID and miniTurbo, which catalyze PL with much greater efficiency than BioID or BioID2, and enable 10-min PL in cells with non-toxic and easily deliverable biotin. Furthermore, TurboID extends biotin-based PL to flies and worms.

Medical subject headings