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GTPγS Assay for Measuring Agonist-Induced Desensitization of Two Human Polymorphic Alpha2B-Adrenoceptor Variants

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Pharmacogenomics in Drug Discovery and Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2547))

Abstract

α2-Adrenergic receptors (ARs) mediate many cellular actions of epinephrine and norepinephrine, including inhibition of their secretion (sympathetic inhibition) from adrenal chromaffin cells. Like many other G protein-coupled receptors (GPCRs), they undergo agonist-dependent phosphorylation and desensitization by GPCR kinases (GRKs), a phenomenon recently shown to play a major role in the sympathetic overdrive that accompanies and aggravates chronic heart failure. A three-glutamic acid deletion polymorphism in the human α2B-AR subtype gene (Glu301–303) causes impaired agonist-promoted receptor phosphorylation and desensitization, resulting in enhanced signaling to inhibition of cholinergic-induced catecholamine secretion in adrenal chromaffin cells. One of the various pharmacological assays that can be used to quantify and quantitatively compare the degrees of agonist-dependent desensitization, i.e., G protein decoupling, of these two polymorphic α2B-AR variants (or of any two GPCRs for that matter) is the guanosine-5′-O-3-thiotriphosphate (GTPγS) assay that can directly quantify heterotrimeric G protein activation.

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Authors and Affiliations

  1. Laboratory for the Study of Neurohormonal Control of the Circulation, Department of Pharmaceutical Sciences, Nova Southeastern University, Fort Lauderdale, FL, USA

    Jordana I. Borges, Alexandra M. Carbone, Natalie Cora, Anastasiya Sizova & Anastasios Lymperopoulos

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  1. Jordana I. Borges
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  2. Alexandra M. Carbone
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  3. Natalie Cora
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  4. Anastasiya Sizova
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  5. Anastasios Lymperopoulos
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Correspondence to Anastasios Lymperopoulos .

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Editors and Affiliations

  1. PharmTao, Santa Clara, CA, USA

    Qing Yan

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Borges, J.I., Carbone, A.M., Cora, N., Sizova, A., Lymperopoulos, A. (2022). GTPγS Assay for Measuring Agonist-Induced Desensitization of Two Human Polymorphic Alpha2B-Adrenoceptor Variants. In: Yan, Q. (eds) Pharmacogenomics in Drug Discovery and Development. Methods in Molecular Biology, vol 2547. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2573-6_12

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  • DOI: https://doi.org/10.1007/978-1-0716-2573-6_12

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2572-9

  • Online ISBN: 978-1-0716-2573-6

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