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Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Technology

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G Protein-Coupled Receptor Screening Assays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2268))

Abstract

Here we describe the stepwise application of bioluminescence resonance energy transfer (BRET)-based conformational receptor biosensors to study GPCR activation in intact cells. This technology can be easily adopted to various plate reader devices and microtiter plate formats. Due to the high sensitivity of these BRET-based receptor biosensors and their ability to quantify simultaneously receptor activation/de-activation kinetics as well as compound efficacy and potency, these optical tools provide the most direct and unbiased approach to monitor GPCR activity in a high-throughput-compatible assay format, representing a novel promising tool for the discovery of potential GPCR therapeutics.

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Acknowledgments

This work was supported by the Federal Ministry of Research (BMBF; 03 V0830), National Institutes of Health (NIH, 0255-8521-4609), SFB/TR166 to M.J.L and by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation – 427840891), Nottingham Research Anne McLaren fellowship to IM.

Competing interests: The authors declare that The University of Würzburg holds a patent on this technology: WO2004057333 A1.

Author information

Authors and Affiliations

  1. Section of Receptor Biology & Signaling, Department of Physiology & Pharmacology, Karolinska Institutet, Stockholm, Sweden

    Hannes Schihada

  2. Institute of Pharmacology and Toxicology, University of Wuerzburg, Wuerzburg, Germany

    Hannes Schihada & Martin J. Lohse

  3. Max-Delbrueck-Center for Molecular Medicine, Berlin, Germany

    Katarina Nemec & Martin J. Lohse

  4. ISAR Bioscience, Planegg, Germany

    Martin J. Lohse

  5. School of Life Sciences, Queen’s Medical Centre, University of Nottingham, Nottingham, UK

    Isabella Maiellaro

Authors
  1. Hannes Schihada
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  2. Katarina Nemec
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  3. Martin J. Lohse
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  4. Isabella Maiellaro
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Corresponding author

Correspondence to Isabella Maiellaro .

Editor information

Editors and Affiliations

  1. INESC Microsistemas e Nanotecnologias, Lisbon, Portugal

    Sofia Aires M. Martins

  2. IBB—Institute for Bioengineering and Biosciences, Lisbon, Portugal

    Duarte Miguel F. Prazeres

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Schihada, H., Nemec, K., Lohse, M.J., Maiellaro, I. (2021). Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Technology. In: Martins, S.A.M., Prazeres, D.M.F. (eds) G Protein-Coupled Receptor Screening Assays. Methods in Molecular Biology, vol 2268. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1221-7_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1221-7_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1220-0

  • Online ISBN: 978-1-0716-1221-7

  • eBook Packages: Springer Protocols

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