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The Rhodopsin-Arrestin-1 Interaction in Bicelles

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Rhodopsin

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1271))

Abstract

G-protein-coupled receptors (GPCRs) are essential mediators of information transfer in eukaryotic cells. Interactions between GPCRs and their binding partners modulate the signaling process. For example, the interaction between GPCR and cognate G protein initiates the signal, while the interaction with cognate arrestin terminates G-protein-mediated signaling. In visual signal transduction, arrestin-1 selectively binds to the phosphorylated light-activated GPCR rhodopsin to terminate rhodopsin signaling. Under physiological conditions, the rhodopsin-arrestin-1 interaction occurs in highly specialized disk membrane in which rhodopsin resides. This membrane is replaced with mimetics when working with purified proteins. While detergents are commonly used as membrane mimetics, most detergents denature arrestin-1, preventing biochemical studies of this interaction. In contrast, bicelles provide a suitable alternative medium. An advantage of bicelles is that they contain lipids, which have been shown to be necessary for normal rhodopsin-arrestin-1 interaction. Here we describe how to reconstitute rhodopsin into bicelles, and how bicelle properties affect the rhodopsin-arrestin-1 interaction.

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Author information

Author notes

  1. Tiandi Zhuang

    Present address: Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, 22908, USA

  2. Min-Kyu Cho

    Present address: Department of Chemistry, New York University, New York, NY, 10003, USA

  3. Tarjani M. Thaker

    Present address: Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, CA, 94158, USA

Authors and Affiliations

  1. Department of Pharmacology, Vanderbilt University Medical Center, 1211 Medical Center Drive, Nashville, TN, 37232-6600, USA

    Qiuyan Chen, Sergey A. Vishnivetskiy, Vsevolod V. Gurevich & T. M. Iverson Ph.D.

  2. Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN, 37232-6600, USA

    Tiandi Zhuang, Min-Kyu Cho, Tarjani M. Thaker, Charles R. Sanders & T. M. Iverson Ph.D.

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  1. Qiuyan Chen
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  2. Sergey A. Vishnivetskiy
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  7. Vsevolod V. Gurevich
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Corresponding author

Correspondence to T. M. Iverson Ph.D. .

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Editors and Affiliations

  1. Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA

    Beata Jastrzebska

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Chen, Q. et al. (2015). The Rhodopsin-Arrestin-1 Interaction in Bicelles. In: Jastrzebska, B. (eds) Rhodopsin. Methods in Molecular Biology, vol 1271. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2330-4_6

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  • DOI: https://doi.org/10.1007/978-1-4939-2330-4_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2329-8

  • Online ISBN: 978-1-4939-2330-4

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