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Recombinant Expression and Purification of Cannabinoid Receptor CB2, a G Protein-Coupled Receptor

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G Protein-Coupled Receptor Screening Assays

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

Abstract

G protein-coupled receptors (GPCR) are integral membrane proteins that regulate multiple cellular processes. To obtain insights into structural properties of GPCR and mechanism of activity, these proteins should be isolated in significant (milligram) quantities, in a pure, homogenous, and stable form. Here we describe the expression and purification of type II human cannabinoid receptor CB2, a class A GPCR, in two different types of expression hosts: in Escherichia coli and in mammalian suspension cell culture Expi293. Our method allows preparation of milligram quantities of the purified receptors suitable for a wide array of downstream applications including high-resolution structural studies and functional assays.

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Acknowledgments

This work was supported by the Intramural Research Program of the National Institute on Alcoholism and Alcohol Abuse, National Institutes of Health. Authors thank Ms. L Zoubak and Mr. K. Hines who contributed to the project and Dr. K. Gawrisch for continuous support.

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Authors and Affiliations

  1. National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD, USA

    Alexei A. Yeliseev

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  1. Alexei A. Yeliseev
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Correspondence to Alexei A. Yeliseev .

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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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Yeliseev, A.A. (2021). Recombinant Expression and Purification of Cannabinoid Receptor CB2, a G Protein-Coupled Receptor. 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_4

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  • DOI: https://doi.org/10.1007/978-1-0716-1221-7_4

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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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