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Interaction of fenoterol stereoisomers with β2-adrenoceptor-Gsα fusion proteins: antagonist and agonist competition binding

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Abstract

The specific interaction between G-protein-coupled receptors and ligand is the starting point for downstream signaling. Fenoterol stereoisomers were successfully used to probe ligand-specific activation (functional selectivity) of the β2-adrenoceptor (β2AR) (Reinartz et al. 2015). In the present study, we extended the pharmacological profile of fenoterol stereoisomers using β2AR-Gsα fusion proteins in agonist and antagonist competition binding assays. Dissociations between binding affinities and effector potencies were found for (R,S′)- and (S,S′)-isomers of 4′-methoxy-1-naphthyl-fenoterol. Our data corroborate former studies on the importance of the aminoalkyl moiety of fenoterol derivatives for functional selectivity.

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Abbreviations

7TMR:

7 transmembrane domain receptor

AC:

Adenylyl cyclase

β2AR:

β2-adrenoceptor

BSA:

Bovine serum albumin

DHA:

Dihydroalprenolol

DMEM:

Dulbecco’s modified Eagle’s medium

EDTA:

Ethylenediaminetetraacetic acid

EPI:

Epinephrine (adrenaline)

Giα:

Inhibitory Gα-protein

Gsα:

Stimulatory Gα-protein

GPCR:

G-protein-coupled receptor

GTPase:

GTP-hydrolyzing activity

ISO:

Isoproterenol

PBS:

Phosphate-buffered saline

PMSF:

Phenylmethanesulfonylfluorid

Sf9:

Clonal isolate of Spodoptera frugiperda cells

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Authors and Affiliations

  1. Institute for Pharmacology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Michael T. Reinartz, Solveig Kälble & Roland Seifert

  2. Laboratory of Clinical Investigation, Biomedical Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD, USA

    Irving W. Wainer

Authors
  1. Michael T. Reinartz
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  2. Solveig Kälble
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  3. Irving W. Wainer
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  4. Roland Seifert
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Correspondence to Roland Seifert.

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Reinartz, M.T., Kälble, S., Wainer, I.W. et al. Interaction of fenoterol stereoisomers with β2-adrenoceptor-Gsα fusion proteins: antagonist and agonist competition binding. Naunyn-Schmiedeberg's Arch Pharmacol 388, 517–524 (2015). https://doi.org/10.1007/s00210-015-1086-5

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  • DOI: https://doi.org/10.1007/s00210-015-1086-5

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