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Ion Channels as Reporters of Membrane Receptor Function: Automated Analysis in Xenopus Oocytes

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Membrane Protein Structure and Function Characterization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1635))

Abstract

G-protein-coupled receptors (GPCR) are the most widely used system of communication used by cells. They sense external signals and translate them into intracellular signals. The information is carried mechanically across the cell membrane, without perturbing its integrity. Agonist binding on the extracellular side causes a change in receptor conformation which propagates to the intracellular side and causes release of activated G-proteins, the first messengers of a variety of signaling cascades.

Permitting access to powerful electrophysiological techniques, ion channels can be employed to monitor precisely the most proximal steps of GPCR signaling, receptor conformational changes, and G-protein release. The former is achieved by physical attachment of a potassium channel to the GPCR to create an Ion-Channel Coupled Receptor (ICCR). The latter is based on the use of G-protein-regulated potassium channels (GIRK). We describe here how these two systems may be used in the Xenopus oocyte heterologous system with a robotic system for increased throughput.

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Acknowledgments

Work in our laboratory is supported by CNRS (Centre National de la Recherche Scientifique), CEA (Commissariat à l’Energie Atomique et aux énergies alternatives), Université Grenoble Alpes, and by grants from the Agence Nationale de la Recherche (VenomPicoScreen project, grant ANR-11-RPIB-022-04) and from the National Institutes of Health (NIH Grant Nr. 5R01EB007047-06). Our laboratory is a member of the French National Laboratory of Excellence “Ion Channel Science and Therapeutics” (LabEX ICST) funded by a network grant from ANR (ANR-11-LABX-0015-01). G.C.M.R. and Z.T. are recipients of doctoral fellowships from LabEX ICST.

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Authors and Affiliations

  1. Institut de Biologie Structurale (IBS), Université Grenoble Alpes, 71 Avenue des Martyrs CS 10090, 38044, Grenoble, Cedex 9, France

    Michel Vivaudou, Zlatomir Todorov & Gina Catalina Reyes-Mejia

  2. CNRS, IBS, LabEx ICST, F-38044, Grenoble, France

    Michel Vivaudou, Zlatomir Todorov & Gina Catalina Reyes-Mejia

  3. CEA, IBS, F-38044, Grenoble, France

    Michel Vivaudou, Zlatomir Todorov & Gina Catalina Reyes-Mejia

  4. Univ. Grenoble Alpes, CNRS, CEA, CNRS, IBS, F-38000, Grenoble, France

    Christophe Moreau

Authors
  1. Michel Vivaudou
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  2. Zlatomir Todorov
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  3. Gina Catalina Reyes-Mejia
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  4. Christophe Moreau
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Corresponding author

Correspondence to Michel Vivaudou .

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Editors and Affiliations

  1. Département de Chimie, CNRS UMR 7203 LBM, Sorbonne Universités—UPMC Univ Paris 06, École Normale Supérieure—PSL Research University, Paris, France

    Jean-Jacques Lacapere

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Vivaudou, M., Todorov, Z., Reyes-Mejia, G.C., Moreau, C. (2017). Ion Channels as Reporters of Membrane Receptor Function: Automated Analysis in Xenopus Oocytes. In: Lacapere, JJ. (eds) Membrane Protein Structure and Function Characterization. Methods in Molecular Biology, vol 1635. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7151-0_15

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  • DOI: https://doi.org/10.1007/978-1-4939-7151-0_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7149-7

  • Online ISBN: 978-1-4939-7151-0

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