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Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) pp 207–226Cite as

Identifying Nuclear Protein–Protein Interactions Using GFP Affinity Purification and SILAC-Based Quantitative Mass Spectrometry

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1188))

Abstract

Many cellular proteins assemble into macromolecular protein complexes. Therefore, identifying protein–protein interactions (PPIs) is essential to gain insight into the function of proteins. Recently established quantitative mass spectrometry-based techniques have significantly improved the unbiased search for PPIs. In this chapter, we describe a single-step GFP affinity purification method combined with SILAC-based quantitative mass spectrometry that can be used to identify nuclear PPIs in mammalian cells.

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Acknowledgments

We thank members of the Vermeulen group and Nina Hubner for critical reading of the manuscript and fruitful discussions. Work in the Vermeulen group is supported by grants from the Netherlands Organisation for Scientific Research (VIDI and Cancer Genomics Centre (CGC)) and the Dutch Cancer Society (KWF).

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Authors and Affiliations

  1. Department of Molecular Cancer Research, University Medical Center Utrecht, 85060, 3508 AB, Utrecht, The Netherlands

    H. Irem Baymaz, Cornelia G. Spruijt & Michiel Vermeulen

Authors
  1. H. Irem Baymaz
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  2. Cornelia G. Spruijt
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  3. Michiel Vermeulen
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Corresponding author

Correspondence to Michiel Vermeulen .

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Editors and Affiliations

  1. Institute of Biology II, University of Freiburg, Freiburg, Germany

    Bettina Warscheid

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Baymaz, H.I., Spruijt, C.G., Vermeulen, M. (2014). Identifying Nuclear Protein–Protein Interactions Using GFP Affinity Purification and SILAC-Based Quantitative Mass Spectrometry. In: Warscheid, B. (eds) Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC). Methods in Molecular Biology, vol 1188. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1142-4_15

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  • DOI: https://doi.org/10.1007/978-1-4939-1142-4_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1141-7

  • Online ISBN: 978-1-4939-1142-4

  • eBook Packages: Springer Protocols

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