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Measuring NLR Oligomerization III: Detection of NLRP3 Complex by Bioluminescence Resonance Energy Transfer

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NLR Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1417))

Abstract

Bioluminescent resonance energy transfer (BRET) is a natural phenomenon resulting from a non-radiative energy transfer between a bioluminescent donor (Renilla luciferase) and a fluorescent protein acceptor. BRET signal is dependent on the distance and the orientation between the donor and the acceptor and could be used to study protein–protein interactions and conformational changes within proteins in real time in living cells. This protocol describes the use of BRET technique to study NLRP3 oligomerization in living cells before and during NLRP3 inflammasome activation.

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Acknowledgements

This work was supported by the European Research Council and instituto salud carlos III-FEDER grants to P.P. F.M.-S. was supported by a Sara Borrell fellowship from the instituto salud carlos III. V.C. was supported by the Institut National de la Santé et de la Recherche Médicale.

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Authors and Affiliations

  1. Inflammation and Experimental Surgery Unit,, Institute for Bio-Health Research of Murcia (IMIB-Arrixaca), Carretera Buenavista, s/n., El Palmar, Murcia, 30120, Spain

    Fátima Martín-Sánchez

  2. Institut de Génomique Fonctionnelle, Labex ICST, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Université Montpellier, Montpellier, France

    Vincent Compan

  3. Institut National de la Santé et de la Recherche Médicale Unité 1191, Montpellier, France

    Vincent Compan

  4. Molecular Inflammation Group, Murcia Biomedical Research Institute (IMIB-Arrixaca), Hospital Virgen de la Arrixaca, Carretera Buenavista, s/n., Murcia, 30120, Spain

    Pablo Pelegrín

Authors
  1. Fátima Martín-Sánchez
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  2. Vincent Compan
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  3. Pablo Pelegrín
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Corresponding author

Correspondence to Fátima Martín-Sánchez .

Editor information

Editors and Affiliations

  1. Department of Morphology Surgery and Experimental Medicine, University of Ferrara, Ferrara, Italy

    Francesco Di Virgilio

  2. Muricia Biomedical Research Institute (IMIB-Arrixaca), Hospital Virgen de la Arrixaca Murcia, Molecular Inflammation Group, Murcia, Spain

    Pablo Pelegrín

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Martín-Sánchez, F., Compan, V., Pelegrín, P. (2016). Measuring NLR Oligomerization III: Detection of NLRP3 Complex by Bioluminescence Resonance Energy Transfer. In: Di Virgilio, F., Pelegrín, P. (eds) NLR Proteins. Methods in Molecular Biology, vol 1417. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3566-6_10

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  • DOI: https://doi.org/10.1007/978-1-4939-3566-6_10

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3564-2

  • Online ISBN: 978-1-4939-3566-6

  • eBook Packages: Springer Protocols

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