MassIVE MSV000082304

Partial Public

Directed evolution of TurboID for efficient proximity labeling in living cells and organisms

Description

Branon TC, Bosch JA, Sanchez AD, Udeshi ND, Svinkina T, Carr SA, Feldman JL, Perrimon N, Ting AY. Nature Biotech 2018. Protein interaction networks and protein compartmentation underlie every signaling process and regulatory mechanism in cells. Recently, proximity labeling (PL) has emerged as a new approach to study the spatial and interaction characteristics of proteins in living cells. However, the two enzymes commonly used for PL come with tradeoffs. BioID is slow, requiring tagging times of 18-24 hours, while APEX peroxidase uses substrates that have limited cell permeability and high toxicity. To address these problems, we used yeast display-based directed evolution to engineer two mutants of biotin ligase, TurboID and miniTurbo, with much greater catalytic efficiency than BioID, and the ability to carry out PL in cells in much shorter time windows (as little as 10 minutes) with non-toxic and easily deliverable biotin. In addition to shortening PL time and increasing PL yield in cell culture, TurboID enabled biotin-based PL in new settings, including Drosophila, and C. elegans. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: TMT ; interaction partners ; proximity labeling

Contact

Principal Investigators:
(in alphabetical order)
Steven A. Carr, Broad Institute of MIT and Harvard, United States
Submitting User: clauser
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