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Observing the Assembly of Protein Complexes in Living Eukaryotic Cells in Super-Resolution Using refSOFI

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Protein Complex Assembly

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1764))

Abstract

Few approaches are currently available that allow the detection of protein-protein interactions (PPIs) in super-resolution, and the observation of the assembly of protein complexes in living cells has been particularly challenging. We developed reconstituted fluorescence-based stochastic optical fluctuation imaging (refSOFI), which is based on bimolecular fluorescence complementation (BiFC) and SOFI, allowing us to detect protein complex assembly 30 min after the induction of complex formation. Here we describe how to use refSOFI to map the assembly of two proteins of interest into a complex within living cells at super-resolution.

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Acknowledgment

This work was supported by NIH (R35 CA197622, R01 DK073368, R01 GM111665, and R01 MH111516 to J.Z.).

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Authors and Affiliations

  1. Department of Chemistry, University of Leuven, Heverlee, Belgium

    Fabian Hertel & Peter Dedecker

  2. Department of Pharmacology, University of California at San Diego, La Jolla, CA, USA

    Gary C. H. Mo & Jin Zhang

Authors
  1. Fabian Hertel
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  2. Gary C. H. Mo
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  3. Peter Dedecker
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  4. Jin Zhang
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Corresponding author

Correspondence to Jin Zhang .

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Editors and Affiliations

  1. MRC Human Genetics Unit, Institute of Genetics & Molecular Medicine, University of Edinburgh, Edinburgh, United Kingdom

    Joseph A. Marsh

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Hertel, F., Mo, G.C.H., Dedecker, P., Zhang, J. (2018). Observing the Assembly of Protein Complexes in Living Eukaryotic Cells in Super-Resolution Using refSOFI. In: Marsh, J. (eds) Protein Complex Assembly. Methods in Molecular Biology, vol 1764. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7759-8_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7759-8_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7758-1

  • Online ISBN: 978-1-4939-7759-8

  • eBook Packages: Springer Protocols

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