Abstract
Proteins often exist and function as part of higher-order complexes or networks. A challenge is to identify the universe of proximal and interacting partners for a given protein. We describe how the high-activity promiscuous biotin ligase called TurboID is fused to the actin-binding peptide LifeAct to label by biotinylation proteins that bind, or are in close proximity, to actin. The rapid enzyme kinetics of TurboID allows the profiles of actin-binding proteins to be compared under different conditions, such as acute disruption of filamentous actin structures with cytochalasin D.
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Acknowledgments
This work was supported by funding to M.F.O. from the Canadian Institutes of Health Research (PJT-169125), Natural Sciences and Engineering Research Council of Canada (RGPIN-2020-05388), and Canada Research Chairs Program (950-231665).
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Joo, E.E., Olson, M.F. (2024). BioID Analysis of Actin-Binding Proteins. In: Nagata, Ki. (eds) Cerebral Cortex Development. Methods in Molecular Biology, vol 2794. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3810-1_9
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DOI: https://doi.org/10.1007/978-1-0716-3810-1_9
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