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NLR Proteins pp 97–113Cite as

Measuring IL-1β Processing by Bioluminescence Sensors II: The iGLuc System

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

Abstract

Inflammasomes are multimeric protein complexes that proteolytically activate caspase-1, which subsequently matures cytokines of the IL-1 family and initiates the induction of pyroptotic cell death. Although this process is central both to pathogen defense and sterile inflammatory processes, there is currently no standard readout available for inflammasome activation which would be suitable for high-throughput applications. We have recently developed a new method for measuring inflammasome activation via the use of a novel proteolytic reporter iGLuc, an IL-1β Gaussia luciferase (iGLuc) fusion protein. Here, we provide detailed protocols for the use of iGLuc in transiently transfected or stably transduced cell lines. Using these protocols, IL-1β maturation as the result of inflammasome activation or other processes can be indirectly measured via the gain of Gaussia luciferase activity of cleaved iGLuc, allowing for rapid inflammasome reconstitution assays and high-throughput screening of inflammasome activity.

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Author information

Authors and Affiliations

  1. Institute of Molecular Medicine, University Hospital, University of Bonn, Sigmund-Freud-Straße 25, Bonn, Germany

    Eva Bartok, Maria Kampes & Veit Hornung

  2. Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Sigmund-Freud-Straße 25, Bonn, Germany

    Eva Bartok

  3. Gene Center and Department of Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen-Str. 25, Munich, Germany

    Veit Hornung

Authors
  1. Eva Bartok
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  2. Maria Kampes
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  3. Veit Hornung
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Corresponding author

Correspondence to Veit Hornung .

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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Bartok, E., Kampes, M., Hornung, V. (2016). Measuring IL-1β Processing by Bioluminescence Sensors II: The iGLuc System. 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_6

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  • DOI: https://doi.org/10.1007/978-1-4939-3566-6_6

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  • 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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