Efficient proximity labeling in living cells and organisms with TurboID
Nature biotechnology, 2018•nature.com
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 …
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 …
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.
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