Abstract
The nuclear envelope (NE) is a critical cellular structure whose constituents and roles in a myriad of cellular processes seem ever expanding. To determine the underlying mechanisms by which the NE constituents participate in various cellular events, it is necessary to understand the nature of their protein-protein associations. BioID (proximity-dependent biotin identification) is a recently established method to generate a history of protein-protein associations as they occur over time in living cells. BioID is based on fusion of a bait protein to a promiscuous biotin ligase. Expression of the BioID fusion protein in a relevant cellular environment enables biotinylation of vicinal and interacting proteins of the bait protein, permitting isolation and identification by conventional biotin-affinity capture and mass-spec analysis. In this way, BioID provides unique capabilities to identify protein-protein associations at the NE. In this chapter we provide a detailed protocol for the application of BioID to the study of NE proteins.
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Acknowledgments
These studies were supported by grants RO1GM102203, RO1GM102486, and RO1EB014869 (to K.J.R.) from the National Institutes of Health and by Sanford Research startup funds (K.J.R.)
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Kim, D.I., Jensen, S.C., Roux, K.J. (2016). Identifying Protein-Protein Associations at the Nuclear Envelope with BioID. In: Shackleton, S., Collas, P., Schirmer, E. (eds) The Nuclear Envelope. Methods in Molecular Biology, vol 1411. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3530-7_8
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DOI: https://doi.org/10.1007/978-1-4939-3530-7_8
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